Yikon Biopsy PGT-A Test (ChromInst Assay) -

20190306
Preimplantation Genetic Screening (PGS)
1. Preimplantation Genetic Screening (PGS) : The practice of evaluating embryos for chromosomal
aneuploidy, the presence of either too many or too few chromosomes, in an effort to select embryos free of
chromosomal aneuploidy for successful uterus implantation and viable pregnancies to deliver normal and
healthy babies.

2. Intended for women

● advanced maternal age
■ over 38 years old
● recurrent pregnancy loss
■ 2 or more times
● recurrent implantation failure
■ 3 or more times
■ failure to implant >= 4 to 6 blastomeres (fragmentation <30%) or >=3 blastocyst (>4BB)
● family history of chromosomal aneuploidy
● carrier of non-homologous Robertsonian translocation (balanced, between acrocentric chromosomes)
● carrier of homologous Robertsonian translocation (balanced, within acrocentric chromosomes)
● carrier of other reciprocal or balanced translocation
ChromInst - Product Features

Fast 9 hour procedure with ~ 1 hour hands-on time

Easy Streamlined procedure for sample prep and data analysis

Accurate WGA technology ensures the uniform and accurate amplification

of the whole genome at the single cell level

High Accurately detect sub-chromosomal CNVs >4Mb

Resolution

Flexible Compatible with both Illumina and Ion Torrent NGS platforms
ChromInst – Embryo Biopsy

1-2 cells 5-10 cells

Day 3 Blastomere Day 5/6 Blastocyst

Biopsy Biopsy (TE Biopsy)
ChromInst Embryo Biopsy – D3 Blastomere Biopsy vs. D5/6 TE Biopsy

Blastomere Embryo Biopsy TE Embryo Biopsy

Fertilization ICSI (highly recommended) ICSI (highly recommended)

Biopsy Timing D3 D5/D6

Biopsy Stage Cleavage stage Blastocyst stage

Biopsy Cells 1-2 single nucleated blastomere 5-10 TE cells out of 100 total TE cells
cell out of 6-8 total blastomere cells (3BB or above) and distant away
with less than 30% fragmentation from ICM
Biopsy Mosaicism Present, more likely Present, less likely

Applicable Embryo with difficulties of in vitro Embryo without difficulties to of in

culture to D5 vitro culture to D5
Embryo development issue Yes Yes(placenta development)
ChromInst Workflow

• Suitable for IVF labs with NGS sequencers

Oocyte Embryo Culture Embryo Biopsy

Result
Reporting
Library Prep
ChromInst Experimental Workflow
ChromInst - Experimental Workflow
ChromInst
a. D3 Blastomere or D5/6 TE Embryo Biopsy Samples
i. D3: 1-2 blastomere cells out of 6-8 total blastomere cells with fragmentation less than 30%
ii. D5/6: a few TE cells (5-10 TE cells typically distant from ICM)
iii. Blank controls: 1uL blank biopsy medium droplet/1uL blank wash droplet after biopsy + 5uL Yikon lysis
buffer; one biopsy blank and one wash blank per IVF batch
b. Cell lysis
i. negative control: 5uL Yikon lysis buffer (contamination in NGS workflow)
ii. positive control: 1uL of 50-100 pg/uL human genomic DNA with known chromosome results + 5uL
Yikon lysis buffer (NGS reagent efficiency)
c. WGA of genomic DNA from lysed cells
d. ChromInst Lib Prep & Next Generation Sequencing
i. Illumina Platform
1. Miniseq/Miseq/Nextseq/Hiseq 2500
ii. Ion Torrent Platform
1. PGM/Ion Proton/S5
e. Detect autosomal and sex chromosome CNV
1. Whole chromosomal level
2. Short and long chromosomal arm level
3. Deletion/duplication (>=10Mb)
4. Mini Deletion / Duplication (>= 4M)
ChromInst Experimental Workflow

● Cell Lysis (Step 1)

○ Biopsied cells stored in 5uL Yikon lysis buffer (buffer solution carried over with the biopsy
sample not to exceed 1uL)
○ Lysis enzyme lyse the biopsied embryo cells to release out genomic DNA
○ Prepare NGS positive/negative control at this step and process IVF controls starting from this
step; sequence both IVF and NGS controls
● Pre-library Preparation (Step 2)
○ WGA amplification of the released genomic DNA from step 1
○ WGA product used as a template for library preparation
● Library Preparation (Step 3)
○ Generate library using WGA products from step 2
■ Introduces adaptor, sequencing read primers, and index primers for genomic sequencing
■ Adaptor ligation, PCR enrichment all in one step
● Magnetic beads library purification (Step 4)
● Library quantification and library pooling (Step 5)
● Post-pooling library quantification for genomic sequencing on Illumina & Ion Torrent platform (Step 6)
● Sequencing preparation and Sequencing (Step 7)
○ Prepare template for sequencing and start sequencing
ChromInst Experimental Workflow

Illumina Ion Torrent

Library fragment size (bp) range 200-1000bp 200-400bp for OT2; 300-
500bp for Ion Chef
Average library fragment size (bp) 300-500bp 300bp for OT2; 400bp for
Ion Chef
Library concentration (ng/ul) range 0.1-60 ng/ul 0.1-20ng/ul

Average library concentration (ng/ul) 20ng/ul 3ng/ul

Magnetic bead purification One step (1X) Two steps: 0.9X, 0.4X for
Ion Chef; 0.75X, 0.15X for
OT2
ChromInst Experimental Workflow - EK kits - Illumina Sequencer

Sequencer Reagent Total raw reads/run Raw read/ sample rxn kit (max # of loading concentration (pM)
(Illumina) (mil) (mil) -deletion 10M samples/run)- deletion 10M
MiniSeq High
Miniseq Output Reagent 22-25 1 24(24) 1.8
Kit (75-cycles)

MiSeq Reagent
Miseq 22-25 1 24(24) 20
Kit v3 (150-cycle)

HiSeq Rapid SBS

Hiseq 2500 250-300 1 24(24) 20
Kit v2 (200 cycles)

NextSeq 500/550
Nextseq 500/550 High Output v2.5 330-400 1 24(24) 2.6
kit (75 cycles)

NextSeq 500/550
Nextseq 500/550 Mid Output v2.5 110-130 1 24(24) 2.6
kit (150 cycles)

Note: Total number of samples to be sequenced includes positive and negative control samples and are calculated based on the total raw read per run for the
selected sequencing instruments and the equal mass pooling using individual library concentration measurement on Qubit after library purification. Sequencing is
SE 55 cycles (insert) + 6 cycles (index). The reaction kit comes in 1 form with 24 unique barcode primers as part of the library prep kit. The conversion factor from
ng/ul on Qubit to nM for the library pool is 3.
ChromInst Experimental Workflow - EK kits - Illumina Sequencer
Sequencer Reagent Total raw reads/ Raw read/ rxn kit (# of rxn kit (max # of loading concentration (pM)
(Illumina) run (mil) sample(mil)-mini- samples/run) - mini- samples/ run)- deletion
deletion 4M deletion 4M 10M
MiniSeq High
Miniseq Output Reagent 22-25 2 24(12) 24(24) 1.8
Kit (75-cycles)
MiSeq Reagent
Miseq Kit v3 (150- 22-25 2 24(12) 24(24) 20
cycle)
HiSeq Rapid
Hiseq 2500 SBS Kit v2 (200 250-300 2 24(24) 24(24) 20
cycles)
NextSeq
500/550 High
Nextseq 500/550 330-400 2 24(24) 24(24) 2.6
Output v2.5 kit
(75 cycles)
NextSeq
500/550 Mid
Nextseq 500/550 110-130 2 24(24) 24(24) 2.6
Output v2.5 kit
(150 cycles)

Note: Total number of samples to be sequenced includes positive and negative control samples and are calculated based on the total raw read per run for the
selected sequencing instruments and the equal mass pooling using individual library concentration measurement on Qubit after library purification. Sequencing is
SE 55 cycles (insert) + 6 cycles (index). The reaction kit comes in 1 form with 24 unique barcode primers as part of the library prep kit. The conversion factor
from ng/ul on Qubit to nM for the library pool is 3.
ChromInst Experimental Workflow - XK kit - Illumina Sequencer

Sequencer Reagent Total raw reads/run Raw read/ sample rxn kit (max # of loading concentration (pM)
(Illumina) (mil) (mil) -deletion 10M samples/run)- deletion 10M
MiniSeq High
Miniseq Output Reagent 22-25 1 24(24)/96(24) 1.8
Kit (75-cycles)

MiSeq Reagent
Miseq 22-25 1 24(24)/96(24) 20
Kit v3 (150-cycle)

HiSeq Rapid SBS

Hiseq 2500 250-300 1 24(96)/96(96) 20
Kit v2 (200 cycles)

NextSeq 500/550
Nextseq 500/550 High Output v2.5 330-400 1 24(96)/96(96) 2.6
kit (75 cycles)

NextSeq 500/550
Nextseq 500/550 Mid Output v2.5 110-130 1 24(96)/96(96) 2.6
kit (150 cycles)

Note: Total number of samples to be sequenced includes positive and negative control samples and are calculated based on the total raw read per run for the
selected sequencing instruments and the equal mass pooling of of individual libraries using individual library concentration measurement on Qubit after library
purification. Sequencing is SE 55 cycles (insert) + 6 cycles (index). The reaction kits come in 2 forms: 24 or 96 library prep reaction kit. The barcodess are
requested separately. The conversion factor from ng/ul on Qubit to nM for the library pool is 3.
ChromInst Experimental Workflow - XK kits - Illumina Sequencer
Sequencer Reagent Total raw reads/ Raw read/ rxn kit (# of rxn kit (max # of loading concentration (pM)
(Illumina) run (mil) sample(mil)-mini- samples/run) - mini- samples/ run)- deletion
deletion 4M deletion 4M 10M
MiniSeq High
Miniseq Output Reagent 22-25 2 24(12)/96(12) 24(24)/96(24) 1.8
Kit (75-cycles)
MiSeq Reagent
Miseq Kit v3 (150- 22-25 2 24(12)/96(12) 24(24)/96(24) 20
cycle)
HiSeq Rapid
Hiseq 2500 SBS Kit v2 (200 250-300 2 24(96)/96(96) 24(96)/96(96) 20
cycles)
NextSeq
500/550 High
Nextseq 500/550 330-400 2 24(96)/96(96) 24(96)/96(96) 2.6
Output v2.5 kit
(75 cycles)
NextSeq
500/550 Mid
Nextseq 500/550 110-130 2 24(48)/96(48) 24(96)/96(96) 2.6
Output v2.5 kit
(150 cycles)

Note: Total number of samples to be sequenced includes positive and negative control samples and are calculated based on the total raw read per run for the
selected sequencing instruments and the equal mass pooling of of individual libraries using individual library concentration measurement on Qubit after library
purification. Sequencing is SE 55 cycles (insert) + 6 cycles (index). The reaction kits come in 2 forms: 24 or 96 library prep reaction kit. The barcodes are
requested separately. The conversion factor from ng/ul on Qubit to nM for the library pool is 3.
 ChromInst Experimental Workflow - EK kits - Thermo Fisher Sequencer
Template rxn kits (max # of loading
Template Generation &
Generation/ Ion Chip Total raw reads/run Raw read/sample # of samples/ run - samples/ run)-p/q arm concentration (pM)
Sequencing Reagent (Ion
Sequencer (Ion Supported (mil) - deletion 10M (mil) - deletion 10M deletion 10M (400k raw reads/
Torrent)
Torrent) sample)

Ion 314 Chip v2

BC/Ion 316 Chip Ion PGM Hi-Q View OT2/Ion 48,96(1)/ 48,96(15)/
OT2/PGM 0-0.55/3-6/4-5.5 1 0-1/2-3 /4-5 25 (5uL)
v2 BC/Ion 318 PGM Hi-Q View Sequencing 48,96(13)
Chip v2 BC

Ion PI Hi-Q OT2 200/Ion PI Hi-Q

OT2/Ion Proton Ion PI Chip v3 60-80 1 60-80 48(48)/96(96) 100 (10uL)
Sequencing 200

Ion 520/Ion 48,96(15)/48,96

OT2/S5 Ion 520/530-OT2 / Ion 540-OT2 3-6/15-20/60-80 1 3-6/15-20/60-80 50 (25uL)
530/Ion 540 (48)/48(48), 96(96)

Ion 520/Ion Ion 520/530-OT2 / Ion 540-OT2 48,96(15)/48,96 50 (25uL)

OT2/S5XL 3-6/15-20/60-80 1 3-6/15-20/60-80
530/Ion 540 (48)/48(48), 96(96)

OT2/S5 Ion 520/Ion 48,96(15)/48,96 50 (25uL)

Ion 520/530-OT2 / Ion 540-OT2 3-6/15-20/60-80 1 3-6/15-20/60-80
GeneStudio 530/Ion 540 (48)/48(48), 96(96)

Note: Total number of samples to be sequenced includes positive and negative controls and and are calculated based on the total raw read per run for the selected sequencing instruments
and the equal mass pooling of individual libraries using individual library concentration measurement on Qubit after library purification. The Ion 520-OT2 and Ion 530-OT2 kit provide
template preparation and sequencing of up to 200 or 400 base-read libraries using OT2 and S5 on Ion 520 and Ion 530 chips. The Ion 540-OT2 kit provides template preparation and
sequencing of up to 200 base-read libraries using OT2 and S5 on Ion 540 chips. The Ion PGM Hi-Q View OT2 and sequencing kit provides template preparation and sequencing of up to
200 or 400 base-read libraries using OT2 and PGM on Ion 314,316, and 318 chips. The Ion PI Hi-Q OT2 and sequencing kit provides template preparation and sequencing of up to 200
base-read libraries using OT2 and Ion Proton on Ion Chip v3. The reaction kits come in two forms: 48 or 96 unique barcode primers as part of the library prep kit. The conversion factor
from ng/ul on Qubit to nM for the library pool is 3.
 ChromInst Experimental Workflow - EK kits - Thermo Fisher Sequencer
Template rxn kit (max # loading
Template Generation & Raw read/ sample
Generation/ Ion Chip Total raw reads /run # of samples/ run- sample/run)-p/q concentration
Sequencing Reagent (Ion (mil)- mini-deletion
Sequencer (Ion Supported (mil) mini-deletion 4M arm(400k raw (pM)
Torrent) 4M
Torrent) read/sample)

Ion 314 Chip v2

BC/Ion 316 Chip Ion PGM Hi-Q View OT2/Ion 48,96(1)/ 48,96(15)/
OT2/PGM 0-0.55/3-6/4-5.5 2 0-1/1-3 /2-3 25 (5uL)
v2 BC/Ion 318 PGM Hi-Q View Sequencing 48,96(13)
Chip v2 BC

Ion PI Hi-Q OT2 200/Ion PI Hi-

OT2/Ion Proton Ion PI Chip v3 60-80 2 30-40 48(48)/96(96) 100 (10uL)
Q Sequencing 200

Ion 520/Ion 48,96(15)/48,96

OT2/S5 Ion 520/530-OT2 / Ion 540-OT2 3-6/15-20/60-80 2 1-3/7-10/30-40 50 (25uL)
530/Ion 540 (48)/48(48), 96(96)

Ion 520/Ion Ion 520/530-OT2 / Ion 540-OT2 48,96(15)/48,96 50 (25uL)

OT2/S5XL 3-6/15-20/60-80 2 1-3/7-10/30-40
530/Ion 540 (48)/48(48), 96(96)

OT2/S5 Ion 520/Ion 48,96(15)/48,96 50 (25uL)

Ion 520/530-OT2 / Ion 540-OT2 3-6/15-20/60-80 2 1-3/7-10/30-40
GeneStudio 530/Ion 540 (48)/48(48), 96(96)

Note: Total number of samples to be sequenced includes positive and negative controls and and are calculated based on the total raw read per run for the selected sequencing instruments
and the equal mass pooling of individual libraries using individual library concentration measurement on Qubit after library purification. The Ion 520-OT2 and Ion 530-OT2 kit provide
template preparation and sequencing of up to 200 or 400 base-read libraries using OT2 and S5 on Ion 520 and Ion 530 chips. The Ion 540-OT2 kit provides template preparation and
sequencing of up to 200 base-read libraries using OT2 and S5 on Ion 540 chips. The Ion PGM Hi-Q View OT2 and sequencing kit provides template preparation and sequencing of up to
200 or 400 base-read libraries using OT2 and PGM on Ion 314,316, and 318 chips. The Ion PI Hi-Q OT2 and sequencing kit provides template preparation and sequencing of up to 200
base-read libraries using OT2 and Ion Proton on Ion 520,530, and 540 chips. The reaction kits come in two forms: 48 or 96 unique barcode primers as part of the library prep kit. The
conversion factor from ng/ul on Qubit to nM for the library pool is 3.
 ChromInst Experimental Workflow - XK kits - Thermo Fisher Sequencer
Template rxn kit (max # of loading
Template Generation &
Generation/ Ion Chip Total raw reads/run Raw read/sample # of samples/ run - samples/ run)-p/q arm concentration
Sequencing Reagent (Ion
Sequencer (Ion Supported (mil) - deletion 10M (mil) - deletion 10M deletion 10M (400k raw reads/ (pM)
Torrent)
Torrent) sample)

Ion 314 Chip v2

BC/Ion 316 Chip Ion PGM Hi-Q View OT2/Ion 48,96(1)/ 48,96(15)/
OT2/PGM 0-0.55/3-6/4-5.5 1 0-1/2-3 /4-5 25 (5uL)
v2 BC/Ion 318 PGM Hi-Q View Sequencing 48,96(13)
Chip v2 BC

Ion PI Hi-Q OT2 200/Ion PI Hi-Q

OT2/Ion Proton Ion PI Chip v3 60-80 1 60-80 48(96)/96(96) 100 (10uL)
Sequencing 200

Ion 520/Ion 48,96(15)/48,96

OT2/S5 Ion 520/530-OT2 / Ion 540-OT2 3-6/15-20/60-80 1 3-6/15-20/60-80 50 (25uL)
530/Ion 540 (48)/48,96(96)

Ion 520/Ion Ion 520/530-OT2 / Ion 540-OT2 48,96(15)/48,96 50 (25uL)

OT2/S5XL 3-6/15-20/60-80 1 3-6/15-20/60-80
530/Ion 540 (48)/48,96(96)

OT2/S5 Ion 520/Ion 48,96(15)/48,96 50 (25uL)

Ion 520/530-OT2 / Ion 540-OT2 3-6/15-20/60-80 1 3-6/15-20/60-80
GeneStudio 530/Ion 540 (48)/48,96(96)

Note: Total number of samples to be sequenced includes positive and negative controls and and are calculated based on the total raw read per run for the selected sequencing instruments
and the equal mass pooling of individual libraries using individual library concentration measurement on Qubit after library purification. The Ion 520-OT2 and Ion 530-OT2 kit provide
template preparation and sequencing of up to 200 or 400 base-read libraries using OT2 and S5 on Ion 520 and Ion 530 chips. The Ion 540-OT2 kit provides template preparation and
sequencing of up to 200 base-read libraries using OT2 and S5 on Ion 540 chips.The Ion PGM Hi-Q View OT2 and sequencing kit provides template preparation and sequencing of up to
200 or 400 base-read libraries using OT2 and PGM on Ion 314,316, and 318 chips. The Ion PI Hi-Q OT2 and sequencing kit provides template preparation and sequencing of up to 200
base-read libraries using OT2 and Ion Proton on Ion Chip v3. There are 48 or 96 unique barcode primers for PGS libraries on Ion Torrent platform. The reaction kits come in two forms: 48
or 96 kits. Barcode can be requested separately. The conversion factor from ng/ul on Qubit to nM for the library pool is 3.
 ChromInst Experimental Workflow - XK kits - Thermo Fisher Sequencer
Template rxn kit (max # loading
Template Generation & Raw read/ sample
Generation/ Ion Chip Total raw reads /run # of samples/ run- sample/run)-p/q concentration
Sequencing Reagent (Ion (mil)- mini-deletion
Sequencer (Ion Supported (mil) mini-deletion 4M arm(400k raw (pM)
Torrent) 4M
Torrent) read/sample)

Ion 314 Chip v2

BC/Ion 316 Chip Ion PGM Hi-Q View OT2/Ion 48,96(1)/ 48,96(15)/
OT2/PGM 0-0.55/3-6/4-5.5 2 0-1/1-3 /2-3 25 (5uL)
v2 BC/Ion 318 PGM Hi-Q View Sequencing 48,96(13)
Chip v2 BC

Ion PI Hi-Q OT2 200/Ion PI Hi-Q

OT2/Ion Proton Ion PI Chip v3 60-80 2 30-40 48(96)/96(96) 100 (10uL)
Sequencing 200

Ion 520/Ion 48,96(15)/48,96

OT2/S5 Ion 520/530-OT2 / Ion 540-OT2 3-6/15-20/60-80 2 1-3/7-10/30-40 50 (25uL)
530/Ion 540 (48)/48,96(96)

Ion 520/Ion Ion 520/530-OT2 / Ion 540-OT2 48,96(15)/48,96 50 (25uL)

OT2/S5XL 3-6/15-20/60-80 2 1-3/7-10/30-40
530/Ion 540 (48)/48,96(96)

OT2/S5 Ion 520/Ion 48,96(15)/48,96 50 (25uL)

Ion 520/530-OT2 / Ion 540-OT2 3-6/15-20/60-80 2 1-3/7-10/30-40
GeneStudio 530/Ion 540 (48)/48,96(96)

Note: Total number of samples to be sequenced includes positive and negative controls and and are calculated based on the total raw read per run for the selected sequencing instruments
and the equal mass pooling of individual libraries using individual library concentration measurement on Qubit after library purification. The Ion 520-OT2 and Ion 530-OT2 kit provide
template preparation and sequencing of up to 200 or 400 base-read libraries using OT2 and S5 on Ion 520 and Ion 530 chips. The Ion 540-OT2 kit provides template preparation and
sequencing of up to 200 base-read libraries using OT2 and S5 on Ion 540 chips. The Ion PGM Hi-Q View OT2 and sequencing kit provides template preparation and sequencing of up to
200 or 400 base-read libraries using OT2 and PGM on Ion 314,316, and 318 chips. The Ion PI Hi-Q OT2 and sequencing kit provides template preparation and sequencing of up to 200
base-read libraries using OT2 and Ion Proton on Ion Chip v3. There are 48 or 96 unique barcode primers for PGS libraries on Ion Torrent platform. The reaction kits come in two forms: 48
or 96 kits.Barcode can be requested separately. The conversion factor from ng/ul on Qubit to nM for the library pool is 3.
 ChromInst Experimental Workflow - EK kits - Thermo Fisher Sequencer
Template rxn kits (max # of loading
Template Generation &
Generation/ Ion Chip Total raw reads/run Raw read/sample # of samples/ run - samples/ run)-p/q arm concentration (pM)
Sequencing Reagent (Ion
Sequencer (Ion Supported (mil) - deletion 10M (mil) - deletion 10M deletion 10M (400k raw reads/
Torrent)
Torrent) sample)

Ion 314 Chip v2

BC/Ion 316 Chip Ion PGM Hi-Q View Chef 400 Kit 48,96(1)/ 48,96(15)/
Chef/PGM 0-0.55/3-6/4-5.5 1 0-1/2-3 /4-5 25 (5uL)
v2 BC/Ion 318 or Ion PGM Hi-Q View Kit 48,96(13)
Chip v2 BC

Chef/Ion Proton Ion PI Chip v3 Ion PI Hi-Q Chef Kit 60-80 1 60-80 48(48)/96(96) 100 (10uL)

Ion 520/Ion 48,96(15)/48,96

Chef/S5 Ion 520/530-Chef / Ion 540-Chef 3-6/15-20/60-80 1 3-6/15-20/60-80 50 (25uL)
530/Ion 540 (48)/48(48), 96(96)

Ion 520/Ion Ion 520/530-Chef / Ion 540-Chef 48,96(15)/48,96 50 (25uL)

Chef/S5XL 3-6/15-20/60-80 1 3-6/15-20/60-80
530/Ion 540 (48)/48(48), 96(96)

Chef/S5 Ion 520/Ion 48,96(15)/48,96 50 (25uL)

Ion 520/530-Chef / Ion 540-Chef 3-6/15-20/60-80 1 3-6/15-20/60-80
GeneStudio 530/Ion 540 (48)/48(48), 96(96)

Note: Total number of samples to be sequenced includes positive and negative controls and and are calculated based on the total raw read per run for the selected sequencing instruments
and the equal mass pooling of individual libraries using individual library concentration measurement on Qubit after library purification. The Ion 520-Chef and Ion 530-Chef kit provide
template preparation and sequencing of up to 200 or 400 base-read libraries using Chef and S5 on Ion 520 and Ion 530 chips. The Ion 540-Chef kit provides template preparation and
sequencing of up to 200 base-read libraries using Chef and S5 on Ion 540 chips. The Ion PGM Hi-Q View Chef 400 kit provides template preparation and sequencing of up to 400 base-read
libraries using Chef and PGM on Ion 314,316, and 318 chips. The Ion PGM Hi-Q View Chef kit provides template preparation and sequencing of up to 200 base-read libraries using Chef
and PGM on Ion 314,316, and 318 chips. The Ion PI Hi-Q Chef kit provides template preparation and sequencing of up to 200 base-read libraries using Chef and Ion Proton on Ion PI Chip
v3. The reaction kits come in two forms: 48 or 96 unique barcode primers as part of the library prep kit. The conversion factor from ng/ul on Qubit to nM for the library pool is 3.
 ChromInst Experimental Workflow - EK kits - Thermo Fisher Sequencer
Template rxn kit (max # loading
Template Generation & Raw read/ sample
Generation/ Ion Chip Total raw reads /run # of samples/ run- sample/run)-p/q concentration (pM)
Sequencing Reagent (Ion (mil)- mini-deletion
Sequencer (Ion Supported (mil) mini-deletion 4M arm(400k raw
Torrent) 4M
Torrent) read/sample)

Ion 314 Chip v2

BC/Ion 316 Chip Ion PGM Hi-Q View Chef 400 Kit 48,96(1)/ 48,96(15)/
Chef/PGM 0-0.55/3-6/4-5.5 2 0-1/1-3 /2-3 25 (5uL)
v2 BC/Ion 318 or Ion PGM Hi-Q View Kit 48,96(13)
Chip v2 BC

Chef/Ion Proton Ion PI Chip v3 Ion PI Hi-Q Chef Kit 60-80 2 30-40 48(48)/96(96) 100 (10uL)

Ion 520/Ion 48,96(15)/48,96

Chef/S5 Ion 520/530-Chef / Ion 540-Chef 3-6/15-20/60-80 2 1-3/7-10/30-40 50 (25uL)
530/Ion 540 (48)/48(48), 96(96)

Ion 520/Ion Ion 520/530-Chef / Ion 540-Chef 48,96(15)/48,96 50 (25uL)

Chef/S5XL 3-6/15-20/60-80 2 1-3/7-10/30-40
530/Ion 540 (48)/48(48), 96(96)

Chef/S5 Ion 520/Ion 48,96(15)/48,96 50 (25uL)

Ion 520/530-Chef / Ion 540-Chef 3-6/15-20/60-80 2 1-3/7-10/30-40
GeneStudio 530/Ion 540 (48)/48(48), 96(96)

Note: Total number of samples to be sequenced includes positive and negative controls and and are calculated based on the total raw read per run for the selected sequencing instruments
and the equal mass pooling of individual libraries using individual library concentration measurement on Qubit after library purification. The Ion 520-Chef and Ion 530-Chef kit provide
template preparation and sequencing of up to 200 or 400 base-read libraries using Chef and S5 on Ion 520 and Ion 530 chips. The Ion 540-Chef kit provides template preparation and
sequencing of up to 200 base-read libraries using Chef and S5 on Ion 540 chips. The Ion PGM Hi-Q View Chef 400 kit provides template preparation and sequencing of up to 400 base-read
libraries using Chef and PGM on Ion 314,316, and 318 chips. The Ion PGM Hi-Q View Chef kit provides template preparation and sequencing of up to 200 base-read libraries using Chef
and PGM on Ion 314,316, and 318 chips. The Ion PI Hi-Q Chef kit provides template preparation and sequencing of up to 200 base-read libraries using Chef and Ion Proton on Ion PI Chip
v3. The reaction kits come in two forms: 48 or 96 unique barcode primers as part of the library prep kit. The conversion factor from ng/ul on Qubit to nM for the library pool is 3.
 ChromInst Experimental Workflow - XK kits - Thermo Fisher Sequencer
Template rxn kit (max # of loading
Template Generation &
Generation/ Ion Chip Total raw reads/run Raw read/sample # of samples/ run - samples/ run)-p/q arm concentration (pM)
Sequencing Reagent (Ion
Sequencer (Ion Supported (mil) - deletion 10M (mil) - deletion 10M deletion 10M (400k raw reads/
Torrent)
Torrent) sample)

Ion 314 Chip v2

BC/Ion 316 Chip Ion PGM Hi-Q View Chef 400 Kit 48,96(1)/ 48,96(15)/
Chef/PGM 0-0.55/3-6/4-5.5 1 0-1/2-3 /4-5 25 (5uL)
v2 BC/Ion 318 or Ion PGM Hi-Q View Kit 48,96(13)
Chip v2 BC

Chef/Ion Proton Ion PI Chip v3 Ion PI Hi-Q Chef Kit 60-80 1 60-80 48(96)/96(96) 100 (10uL)

Ion 520/Ion 48,96(15)/48,96

Chef/S5 Ion 520/530-Chef / Ion 540-Chef 3-6/15-20/60-80 1 3-6/15-20/60-80 50 (25uL)
530/Ion 540 (48)/48,96(96)

Ion 520/Ion Ion 520/530-Chef / Ion 540-Chef 48,96(15)/48,96 50 (25uL)

Chef/S5XL 3-6/15-20/60-80 1 3-6/15-20/60-80
530/Ion 540 (48)/48,96(96)

Chef/S5 Ion 520/Ion 48,96(15)/48,96 50 (25uL)

Ion 520/530-Chef / Ion 540-Chef 3-6/15-20/60-80 1 3-6/15-20/60-80
GeneStudio 530/Ion 540 (48)/48,96(96)

Note: Total number of samples to be sequenced includes positive and negative controls and and are calculated based on the total raw read per run for the selected sequencing instruments
and the equal mass pooling of individual libraries using individual library concentration measurement on Qubit after library purification. The Ion 520-Chef and Ion 530-Chef kit provide
template preparation and sequencing of up to 200 or 400 base-read libraries using Chef and S5 on Ion 520 and Ion 530 chips. The Ion 540-Chef kit provides template preparation and
sequencing of up to 200 base-read libraries using Chef and S5 on Ion 540 chips.The Ion PGM Hi-Q View Chef 400 kit provides template preparation and sequencing of up to 400 base-read
libraries using Chef and PGM on Ion 314,316, and 318 chips. The Ion PGM Hi-Q View Chef kit provides template preparation and sequencing of up to 200 base-read libraries using Chef
and PGM on Ion 314,316, and 318 chips. The Ion PI Hi-Q Chef kit provides template preparation and sequencing of up to 200 base-read libraries using Chef and Ion Proton on Ion PI Chip
v3. There are 48 or 96 unique barcode primers for PGS libraries on Ion Torrent platform. The reaction kits come in two forms: 48 or 96 kits.Barcode can be requested separately.The
conversion factor from ng/ul on Qubit to nM for the library pool is 3.
 ChromInst Experimental Workflow - XK kits - Thermo Fisher Sequencer
Template rxn kit (max # loading
Template Generation & Raw read/ sample
Generation/ Ion Chip Total raw reads /run # of samples/ run- sample/run)-p/q concentration (pM)
Sequencing Reagent (Ion (mil)- mini-deletion
Sequencer (Ion Supported (mil) mini-deletion 4M arm(400k raw
Torrent) 4M
Torrent) read/sample)

Ion 314 Chip v2

BC/Ion 316 Chip Ion PGM Hi-Q View Chef 400 Kit 48,96(1)/ 48,96(15)/
Chef/PGM 0-0.55/3-6/4-5.5 2 0-1/1-3 /2-3 25 (5uL)
v2 BC/Ion 318 or Ion PGM Hi-Q View Kit 48,96(13)
Chip v2 BC

Chef/Ion Proton Ion PI Chip v3 Ion PI Hi-Q Chef Kit 60-80 2 30-40 48(96)/96(96) 100 (10uL)

Ion 520/Ion 48,96(15)/48,96

Chef/S5 Ion 520/530-Chef / Ion 540-Chef 3-6/15-20/60-80 2 1-3/7-10/30-40 50 (25uL)
530/Ion 540 (48)/48,96(96)

Ion 520/Ion Ion 520/530-Chef / Ion 540-Chef 48,96(15)/48,96 50 (25uL)

Chef/S5XL 3-6/15-20/60-80 2 1-3/7-10/30-40
530/Ion 540 (48)/48,96(96)

Chef/S5 Ion 520/Ion 48,96(15)/48,96 50 (25uL)

Ion 520/530-Chef / Ion 540-Chef 3-6/15-20/60-80 2 1-3/7-10/30-40
GeneStudio 530/Ion 540 (48)/48,96(96)

Note: Total number of samples to be sequenced includes positive and negative controls and and are calculated based on the total raw read per run for the selected sequencing instruments
and the equal mass pooling of individual libraries using individual library concentration measurement on Qubit after library purification. The Ion 520-Chef and Ion 530-Chef kit provide
template preparation and sequencing of up to 200 or 400 base-read libraries using Chef and S5 on Ion 520 and Ion 530 chips. The Ion 540-Chef kit provides template preparation and
sequencing of up to 200 base-read libraries using Chef and S5 on Ion 540 chips. The Ion PGM Hi-Q View Chef 400 kit provides template preparation and sequencing of up to 400 base-read
libraries using Chef and PGM on Ion 314,316, and 318 chips. The Ion PGM Hi-Q View Chef kit provides template preparation and sequencing of up to 200 base-read libraries using Chef
and PGM on Ion 314,316, and 318 chips. The Ion PI Hi-Q Chef kit provides template preparation and sequencing of up to 200 base-read libraries using Chef and Ion Proton on Ion PI Chip
v3. There are 48 or 96 unique barcode primers for PGS libraries on Ion Torrent platform. The reaction kits come in two forms: 48 or 96 kits.Barcode can be requested separately. The
conversion factor from ng/ul on Qubit to nM for the library pool is 3.
ChromInst Experimental Workflow - Uniformity Add-On (Illumina) - EK Kits
Sequencer Reagent Total raw Raw read/ sample rxn kits (# of rxn kits (max # of Yikon loading concentration
(Illumina) reads/run (mil) (mil) -deletion 10M samples/run)- deletion samples/run) - p/q arm (pM)
10M (400K raw reads/ sample)
MiniSeq High
Miniseq Output Reagent 22-25 1 24(24) 24(24) 1.4
Kit (75-cycles)
MiSeq Reagent
Miseq Kit v3 (150- 22-25 1 24(24) 24(24) 20
cycle)
HiSeq Rapid
Hiseq 2500 SBS Kit v2 (200 250-300 1 24(24) 24(24) 20
cycles)
NextSeq
500/550 High
Nextseq 500/550 330-400 1 24(24) 24(24) 2.6
Output v2.5 kit
(75 cycles)
NextSeq
500/550 Mid
Nextseq 500/550 110-130 1 24(24) 24(24) 2.6
Output v2.5 kit
(150 cycles)

Note: Total number of samples to be sequenced includes positive and negative control samples and are calculated based on the total raw read per run for
the selected sequencing instruments and the equal mass pooling using individual library concentration measurement on Qubit after library purification.
Sequencing is SE 55 cycles (insert) + 6 cycles (index). Each uniformity kit can support two 24 rxn EK kits on two sequencing runs.The conversion factor
from ng/ul on Qubit to nM for the library pool is 11.
ChromInst Experimental Workflow - Uniformity Add-On (Illumina) - EK kits
Sequencer Reagent Total raw Raw read/ rxn kit (# of rxn kit (max # of Yikon loading concentration (pM)
(Illumina) reads/run (mil) sample(mil)-mini- samples/run) - mini- samples/run)- p/q arm
deletion 4M deletion 4M level (400k raw
reads/sample
MiniSeq High
Miniseq Output Reagent 22-25 2 24(12) 24(24) 1.4
Kit (75-cycles)
MiSeq Reagent
Miseq Kit v3 (150- 22-25 2 24(12) 24(24) 20
cycle)
HiSeq Rapid
Hiseq 2500 SBS Kit v2 (200 250-300 2 24(24) 24(24) 20
cycles)
NextSeq
500/550 High
Nextseq 500/550 330-400 2 24(24) 24(24) 2.6
Output v2.5 kit
(75 cycles)
NextSeq
500/550 Mid
Nextseq 500/550 110-130 2 24(24) 24(24) 2.6
Output v2.5 kit
(150 cycles)

Note: Total number of samples to be sequenced includes positive and negative control samples and are calculated based on the total raw read per run for the
selected sequencing instruments and the equal mass pooling using individual library concentration measurement on Qubit after library purification. Sequencing is
SE 55 cycles (insert) + 6 cycles (index). Each uniformity kit can support two 24 rxn EK kits on two sequencing runs. The conversion factor from ng/ul on Qubit to
nM for the library pool is 11.
 ChromInst Experimental Workflow - Uniformity Add-On (Illumina)- XK kits
Sequencer Reagent Total raw Raw read/ sample rxn kit (# of rxn kit (max # of loading concentration (pM)
(Illumina) reads/run (mil) (mil) -deletion 10M samples/run)- deletion samples/run) - p/q arm
10M (400K raw reads/ sample)
MiniSeq High
Miniseq Output Reagent 22-25 1 24(24)/96(24) 24(48)/96(48) 1.8
Kit (75-cycles)
MiSeq Reagent
Miseq Kit v3 (150- 22-25 1 24(24)/96(24) 24(48)/96(48) 20
cycle)
HiSeq Rapid
Hiseq 2500 SBS Kit v2 (200 250-300 1 24(96)/96(96) 24(96)/96(96) 20
cycles)
NextSeq
500/550 High
Nextseq 500/550 330-400 1 24(96)/96(96) 24(96)/96(96) 2.6
Output v2.5 kit
(75 cycles)
NextSeq
500/550 Mid
Nextseq 500/550 110-130 1 24(96)/96(96) 24(96)/96(96) 2.6
Output v2.5 kit
(150 cycles)

Note: Total number of samples to be sequenced includes positive and negative control samples and are calculated based on the total raw read per run for the selected
sequencing instruments and the equal mass pooling of of individual libraries using individual library concentration measurement on Qubit after library purification.
Sequencing is SE 55 cycles (insert) + 6 cycles (index). The reaction kits come in 2 forms: 24 or 96 library prep reaction kit. The barcodes are requested separately.
Each uniformity kit can support 48 reactions using two 24 rxn XK kits or one 96 rxn XK kit with 48 unique barcodes. The conversion factor from ng/ul on Qubit to nM for
the library pool is 11.
ChromInst Experimental Workflow - Uniformity Add-On (Illumina) - XK kits
Sequencer Reagent Total raw Raw read/ rxn kit (# of rxn kit (max # of Yikon loading concentration (pM)
(Illumina) reads/run (mil) sample(mil)-mini- samples/run) - mini- samples/run)- p/q arm
deletion 4M deletion 4M level (400k raw
reads/sample
MiniSeq High
Miniseq Output Reagent 22-25 2 24(12)/96(12) 24(48)/96(48) 1.4
Kit (75-cycles)
MiSeq Reagent
Miseq Kit v3 (150- 22-25 2 24(12)/96(12) 24(48)/96(48) 20
cycle)
HiSeq Rapid
Hiseq 2500 SBS Kit v2 (200 250-300 2 24(96)/96(96) 24(96)/96(96) 20
cycles)
NextSeq
500/550 High
Nextseq 500/550 330-400 2 24(96)/96(96) 24(96)/96(96) 2.6
Output v2.5 kit
(75 cycles)
NextSeq
500/550 Mid
Nextseq 500/550 110-130 2 24(48)/96(48) 24(96)/96(96) 2.6
Output v2.5 kit
(150 cycles)

Note: Total number of samples to be sequenced includes positive and negative control samples and are calculated based on the total raw read per run for the selected
sequencing instruments and the equal mass pooling of of individual libraries using individual library concentration measurement on Qubit after library purification.
Sequencing is SE 55 cycles (insert) + 6 cycles (index). The reaction kits come in 2 forms: 24 or 96 library prep reaction kit. The barcodes are requested separately.
Each uniformity kit can support 48 reactions using two 24 rxn XK kits or one 96 rxn XK kit using 48 unique barcodes. The conversion factor from ng/ul on Qubit to nM
for the library pool is 11.
ChromInst Experimental Workflow - Uniformity Product (Illumina)

Illumina

Individual uniformity product fragment size (bp) range 200-1000bp

Average individual uniformity product fragment size (bp) 300-500bp

Individual uniformity product concentration (ng/ul) range 19-32 ng/ul

Average individual uniformity product concentration (ng/ul) 25ng/ul

Magnetic bead purification One step (1X)

Shipping ChromInst Libraries at Room Temperature

● Inst RT Shipping Kit and Protocol for ChromInst libraries:

- 5ul pre-added preservatives allowing shipping ChromInst libraries at room temperature to Yikon
China for library purification, sequencing, and data analysis
- ChromInst libraries made according to ChromInst Library Prep Protocol from
cell lysis through library preparation (before library purification)
- Feasible option if a lab has no sequencing capacity or if a lab wants to use test samples for trials
for the ChromInst test
ChromInst - Data Analysis & Report Generation
ChromInst ChromGo - Data Analysis & Report Generation

● In-house software for data analysis & report generation

● No Bioinformatics expertise needed
○ Easy-to-use and user-friendly interface with minimal hands-on time
○ Onsite software provided
● Flexibility and adaptability on sequencing platform
○ Can support both Illumina (FASTQ) and Ion Torrent (FASTQ and BAM)
● Customized and automatic procedures
○ Data analysis and report at different resolution
■ Whole chromosome
■ Long/short chromosome arm (p/q arm)
■ Deletion/Duplication >= 10Mb
■ Mini Deletion / Duplication >= 4M
○ Enable chromosome mosaicism and gender information reporting
● Powerful and intuitive data visualization
○ CNV plot at whole genome level and at individual chromosome level
● Support both local and cloud-based server
○ Analyze sequencing data and generate report on ChromGo with locally installed servers or by
connecting to cloud-based servers
 ChromInst ChromGo - Data QC - Valid Reads

Valid Reads
Data Resolution
[0,50K] (50K,100K] (100K,200K] (200K,500K] (500K,1M] (1M,3M] (3M,+∞)

Whole Chromosome FAIL WARNING PASS PASS PASS PASS PASS

Whole Arm (p/q arm) FAIL WARNING WARNING PASS PASS PASS PASS

Deletion / Duplication (10M) FAIL WARNING WARNING WARNING PASS PASS PASS

Mini Deletion / Duplication (4M) FAIL WARNING WARNING WARNING WARNING PASS PASS

Recommended minimum raw reads/ recommended minimum valid reads (CNV accuracies are affected with lower valid read numbers)
● Whole Chromosome: 200K/100K
● Whole Arm (p/q arm): 400K/200K
● Deletion/Duplication (10M): 1M/500K
● Mini Deletion/Duplication (4M): 2M/1M
ChromInst ChromGo - Data QC - Valid Reads QC Score
ChromInst ChromGo - Data QC - Valid Read GC Contents

Valid Read GC Contents (%)

Data Resolution
[0,36] (36,38] (38,43) [43,48) [48,+∞)

Whole Chromosome WARNING INFO PASS INFO WARNING

Whole Arm (p/q arm) WARNING INFO PASS INFO WARNING

Deletion / Duplication (10M) WARNING INFO PASS INFO WARNING

Mini Deletion / Duplication (4M) WARNING INFO PASS INFO WARNING

ChromInst ChromGo - Data QC - Valid Read GC Contents QC Score
ChromInst ChromGo - Data QC - CNV CV

CV(1000K bin size)

Resolution
[0,0.12] (0.12,0.20] (0.2,0.25] (0.25,+∞) NA

Whole Chromosome PASS INFO WARNING FAIL FAIL

Whole Arm (p/q arm) PASS INFO WARNING FAIL FAIL

Deletion / Duplication (10M) PASS INFO WARNING FAIL FAIL

Mini Deletion / Duplication (4M) PASS INFO WARNING FAIL FAIL

ChromInst ChromGo - Data QC - CNV CV QC Score
ChromInst ChromGo - Data QC Score

● Individual valid reads QC score, valid read GC content (%) score, CNV CV QC score
○ 0: Pass
○ 1: Info
○ 2: Warning
○ 3: Serious Warning
○ 4: Fail
● Overall QC Score
○ Individual valid reads QC score + valid read GC content QC score + CNV CV QC score
○ Data QC conclusion based overall QC score
■ Fail: 4 and above
■ Warning: 2 and 3
■ Info: 1
■ Pass: 0
ChromInst ChromGo – Ploidy and Mosaicism Reporting - Mosaicism Included

Report Mosaicism

Aneuploidy Mosaicism

Resolution

Chr 13,16,18,21 Chr 19 Others

Ratio Length

Ratio Length Ratio Length Ratio Length

[0.7,1.3)
Whole Chromosome Whole Chromosome [30%,70%) Whole Chromosome [50%,70%) Whole Chromosome [40%,70%) Whole Chromosome
[2.7,3.3)

[0.7,1.3) Whole Arm/Whole Whole Arm/Whole Whole Arm/Whole Whole Arm /Whole
Whole Arm (p/q arm) [30%,70%) [50%,70%) [40%,70%)
[2.7,3.3) Chromosome Chromosome Chromosome Chromosome

[0.7,1.3) 10M/Whole Arm /Whole 30M/Whole Arm /Whole Whole Arm/Whole 40M /Whole Arm
Deletion / Duplication (10M) [30%,70%) [50%,70%) [40%,70%)
[2.7,3.3) Chromosome Chromosome Chromosome /Whole Chromosome

[0.7,1.3) 4M/10M/Whole Arm 30M/Whole Arm /Whole Whole Arm/Whole 40M/Whole Arm /Whole
Mini Deletion / Duplication (4M) [30%,70%) [50%,70%) [40%,70%)
[2.7,3.3) /Whole Chromosome Chromosome Chromosome Chromosome
ChromInst ChromGo – Ploidy Reporting - Mosaicism Not Selected

No Report Mosaicism

Resolution Length Aneuploidy

Copy Number (CN)

Whole Chromosome Whole Chromosome Round (>=0.5 CN 1 up; <0.5 CN 1 down)

Whole Arm (>30M)/Whole

Whole Arm (p/q arm) Round (>=0.5 CN 1 up; <0.5 1 CN down)
Chromosome
10M/Whole Arm (>30M)/Whole
Deletion / Duplication (10M) Round (>=0.5 CN 1 up; <0.5 CN 1 down)
Chromosome
4M/10M/ Whole Arm
Mini Deletion / Duplication (4M) Round (>=0.5 CN 1 up; <0.5 CN 1 down)
(>30M)/Whole Chromosome
ChromInst ChromGo - Hardware Requirements
ChromSwift Experimental and Data Analysis Workflow

• Suitable for IVF labs without NGS sequencers

Sample Quality Control Report Delivery within

Biopsy Preparation Shipping RT
~15 min 7-10 business days
~ 2.5hr
ChromSwift Product Features

Fast 2.5-hour procedure with ~ 5 min hands-on time

Easy Streamlined 2-step procedure

Accurate WGA technology ensures the uniform and accurate amplification

of the whole genome at the single cell level

Safer
Room temperature shipment of amplified biopsy samples
Shipment

Lower Cost No need for dry ice shipment

ChromSwift - Experimental Workflow

● Cell Lysis (Step 1)

○ Biopsied cells stored in Yikon-supplied lysis buffer
○ Lysis enzyme lyse the biopsied embryo cells to release out genomic DNA
○ Negative control: 4.5uL Yikon lysis buffer
○ Positive control: 1uL of 50 pg/uL human genomic DNA added to 4.5uL Yikon lysis buffer
○ Control samples start from this step; sequencing needed for the control samples
● Amplification (Step 2)
○ WGA of the released genomic DNA from step 1
● Library Preparation (Step 3)
○ This step and subsequent steps are not performed by customers
○ The WGA product from step 2 is shipped to Yikon at room temperature for library preparation
and genomic sequencing
○ Data analysis and report generation performed by Yikon
■ Yikon provides data analysis results and reports to the customers
■ ChromGo software is not needed by the customers
ChromSwift - Experimental Workflow
Shipping ChromInst Libraries at Room Temperature

● Inst Product RT Shipping Kit and Protocol for ChromInst libraries:

- 5ul pre-added preservatives allowing shipping ChromInst libraries at room temperature to Yikon
China for library purification, sequencing, and data analysis
- ChromInstInst libraries made according to ChromInst Library Prep Protocol from cell lysis
through library preparation (before library purification)
- Feasible option if a lab has no sequencing capacity or if a lab wants to use test samples for trials
Validation of ChromInst - ChromInst vs aCGH Assay

Previous aCGH
Assay on initial
30 Blastocyst with TE Biopsy
aneuploidy previously PGS Result
detected by aCGH
assay on initial TE
biopsy (3-6 cells) TE Re-Biopsy
after Thawing ChromInst
(3-5 cells)

Key Performance Parameters ChromInst vs aCGH

Sensitivity 91.11%
Specificity 99.85%
Positive predictive value (PPV) 97.62%
Negative predictive value (NPV) 99.41%
Validation of ChromInst - ChromInst vs SNP Assay

TE Re-Biopsy
after Thawing SNP Assay
(3-5 cells)
30 Blastocyst with
aneuploidy previously PGS Result
detected by aCGH
assay on initial TE
biopsy (3-6 cells) TE Re-Biopsy
after Thawing ChromInst
(3-5 cells)

Key Performance Parameters ChromInst vs SNP

Sensitivity 95.35%
Specificity 99.85%
Positive predictive value (PPV) 97.62%
Negative predictive value (NPV) 99.71%
Example of ChromInst Results on ChromGo

47,XN,+4

Multiple Chromosomal
Aneuploidy

46,XN

46,XN,+6q(q24.1→qter)

46,XN,-5 (mos*)
ChromInst Summary

● High accuracy in screening out normal chromosomes and chromosomes with aneuploidy
○ Good concordance between ChromInst and SNP
■ Specificity and NPV > 95% & Sensitivity and PPV > 95%
○ Good concordance between ChromInst and aCGH
■ Specificity and NPV > 95% & Sensitivity >90% and PPV > 95%
○ All 30 blastocyst embryos correctly identifies as aneuploidy embryos among the three methods
■ 100% aneuploidy detection
● Much lower chromosome mosaicism or FP in D5 blastocyst biopsy than D3 cleavage-stage biopsy
○ D5 blastocyst biopsy (30 blastocyst embryos)
■ 26 embryos with identical aneuploidy results among all three methods (86.67%)
○ D3 cleavage-stage biopsy (23 blastocyst embryos)
■ 8 embryos with identical aneuploidy results among all three methods (34.78%)
● Benefits of ChromInst WGA Technology
○ Good amplification uniformity, high genome coverage, low allele dropout rate, high sensitivity
● Benefits of ChromInst
○ Fast, easy, accurate, high resolution, flexible
Publication of ChromInst

Huang J et al. Fertil & steril, 2014; 102: 1685-1691

Huang J et a.l, Fertil Steril. 2016 Jun;105(6):1532-6.
More Publication on Yikon ChromInst

•Genome-Wide Detection of Single-Nucleotide and Copy-Number Variations of a Single Human Cell. Science, 2012,
2012 338, 1622.
• Probing Meiotic Recombination and Aneuploidy of Single Sperm Cells by Whole-Genome Sequencing.
2012 Science, 2012, 338(6114, 1627-1630)

• Genome Analyses of Single Human Oocytes. Cell, 2013, 155: 1492-1506

2013
• Reproducible Copy Number Variation Patterns Among Single Circulating Tumor Cells of Lung Cancer Patients. PNAS,
2013 2013, 110(52): 21083-21088
• Validation of multiple annealing and looping-based amplification cycle sequencing for 24-chromosome aneuploidy
2014 screening of cleavage-stage embryos. Fertility and Sterility, 2014, 102: 1685-1691
• Single-Cell Whole-Genome Amplification and Sequencing: Methodology and Applications. Annu. Rev. Genomics Hum.
2015 Genet., 2015, 16: 16.1-16.24
• Live Births after Simultaneous Avoidance of Monogenic Diseases and Chromosome Abnormality by Next-Generation
2015 Sequencing with Linkage Analyses . PNAS, 2015, 112(52):15964-9
• A New Next-generation Sequencing-based Assay for Concurrent Preimplantation Genetic Diagnosis of Charcot-Marie-
2016 Tooth Disease Type A1 and Aneuploidy Screening. JGG, 2016, Available online 21 January 2016
• Validation of a next-generation sequencing-based protocol for 24-chromosome aneuploidy screening of blastocysts.
2016 Fertility and Sterility, 2016
Thank You!