BI-113 Prac�cal Study Guide

This study guide is meant to help you prepare for the prac�cal. This should not be the only thing you
study. Please review the handouts and slideshows from each lab as well. Also review the
recommended content in the prac�cal review slideshow we covered in class and was shared with you
in a blackboard announcement. The slideshows can be found in the lab presenta�on and notes folder
on blackboard. The handouts can be found in the weekly folders under the Lab Ac�vi�es folder on Black
board.

Pipe�ng
Unit Conversion

1ml=1,000µl

1L= 1,000ml

*Pay attention to units! They will be used interchangeably but can have a big impact on the result if
the wrong unit is used. This can lead to volume errors if units aren’t interpreted and converted
properly.
Solu�ons Prac�ce Problems

1. You want to make 500 ml of 0.18 M NaCl (MW = 58.44). What mass of NaCl do you
use? Show your work. A molarity problem, so make sure to express all units fully.

2. How many grams of potassium carbonate, K2CO3 (MW: 138), are needed to make 250
mL of a 2.5 M solution?
 3. How many liters of 4.0 M solution can be made using 125 grams of lithium bromide,
LiBr (MW: 86.8)?

4. How many grams of copper (II) fluoride, CuF2 (MW: 101.53), are needed to make 6.7 liters
of a 1.2 M solution?
Dilu�ons Prac�ce Problems

Dilutions Formula

CsVs = CdVd

Cs= Concentration Stock Solution

Vs= Volume Stock Solution
Cd= Concentration Diluted Solution
Vd= Volume Diluted Solution

1. If I have 340 mL of a 0.5 M NaBr solution, what will the concentration be if I add 560
mL more water to it?

*Rember that dilutions questions you can check your work by confirming that each side of the
equation is equal to the other.
0.5 x 340 = 170
0.1888889 x 900 = 170
This is the best way to confirm if you put the variables in the correct places in the experiment.
2. If I dilute 250 mL of 0.10 M lithium acetate solution to a volume of 750 mL, what will the
concentration of this solution be?

2. If I leave 750 mL of 0.50 M sodium chloride solution uncovered on a windowsill and 150
mL of the solvent evaporates, what will the new concentration of the sodium chloride
 solution be?
*Think about what happens to Vd if when there is evaporation. In this style of question Vd
will be lower the Vs.

3. To what volume would I need to add water to the evaporated solution in problem 3 to get
a solution with a concentration of 0.25 M?
*In this question you are figuring out what Vd would be to reduce the concentration you
solved for in the question above to 0.25M.

5. You do a dilution by combining 100 ml volume of MgCl plus 700 ml unit volumes of RO
water. What is the dilution factor, i.e, how many more times dilute is it than the original
concentration?

6. You have a 100 ml stock solution of 100 mg/ml ampicillin in deionized water. You want to
make 30 ml of 25 mg/ml ampicillin in deionized water. How much ampiciliin stock and how
much deionized water are needed to make this?

*For this type of question, do not be confused that the concentration is in mg/ml instead of
Molarity (M) like you are used to. Treat it the same way and use the same formula.
* CAREFULLY READ THIS QUESTION AND PICK OUT THE RELEVANT PIECES OF
INFORMATION THAT NEED TO BE PLUGGED INTO THE FORMULA.
*Focus on the last sentence and what 2 pieces of information it is asking you for.
*Hint: The volume of stock solution mentioned in the question is not what will get plugged
into the equation as Cs.
* Remember you can check your answer by verifying if the two sides of the equation are equal
to each other.
Beer Me

• Review the handwriten notes on the following pages, that were atached to the email with
your lab report grade.
• Focus on understanding how the experiment works, which means how manipula�ng your
variable (mashing temp or pH) impacts the ac�vity of alpha and beta amalyase.
• Why does increased or decreased alpha and beta amalyase ac�vity result in:
o More/less reducing carbohydrates at 50 min of mashing?
o A greater/lesser amount of small fermentable sugars at 50 min of mashing?
o More/less alcohol post fermenta�on?
o More/less reducing carbohydrates remaining post fermenta�on? What does more
reducing carbohydrates le� a�er mashing indicate?
• Understand what the change in mass of the flask contents from pre fermenta�on to post
fermenta�on means, what is leaving the flask, and how is this related to alcohol produc�on?
o The weight lost from the flask during fermenta�on is due to CO2, a byproduct of
fermenta�on, alcohol is the other byproduct. The number of molecules of CO2 and
alcohol produced during fermenta�on is equal. Review the formula in the powerpoint
and week 3 handout.
o The mass of CO2 released from the flask is used to solve for the mass of alcohol
produced during fermenta�on using the molar weights of each compound, as well as
the rela�onship between the amount of CO2 and alcohol.
pGLO

• Understand the set up of the experimental system.

o We studied E.Coli a type of bacteria, that contain the pGLO plasmid, and the pGLO
plasmid was mutated with the TN5 transposon. Be able to define each component of the
system and how they relate to each other.
o Be able to interpret the phenotypes we studied in lab (Kanamycin Resistance, and
Ampicillin Resistance, and GFP expression.) This could be interpre�ng where TN5
inserted based on a table of phenotypes, from looking at plates of bacteria, or plasmid
maps.
o Understand what each phenotype means in terms of where TN5 inserted. If TN5 inserts
in a gene or its promoter (GFP/AmpR) the func�on of that gene will be lost, which
changes the phenotype. If TN5 inserted into a non coding region there will be no impact
to the phenotype other than the E.Coli Clone will gain the ability to resist the an�bio�c
Kanamycin.
o Understand the structure of the pGLO plasmid and be able to interpret a plasmid map.
Be familiar with key terms/concepts such as coding region, non coding region,
recogni�on site, restric�on enzyme, restric�on enzyme fragment, restric�on enzyme
diges�on.
o Understand how we used restric�on enzymes to cut the pGLO plasmid into fragments,
and why this was done. The length of each fragment in an unmutated plasmid is known.
TN5 is 1230 base pairs long. The fragment that TN5 inserts in, will increase in length by
1230 base pairs, while the other 4 fragments will remain the same length. By measuring
the length of each fragment in a mutated pGLO and comparing to an unmutated plasmid
we can determine which of the 5 fragments pGLO inserted in.
o The length of each fragment is evaluated using gel electrophoresis.