DNA Extraction
Before you start the experiment, read the following information which explains what
you will be observing in this extraction process. It explains what actually happens:
The cells are made up of strong cell walls, fatty
cell membranes and contain a nucleus with DNA
inside and lots of proteins
The high temperature breaks down the cell
walls exposing the cell membranes.
The detergent in the washing up liquid dissolves the
fatty cell membranes.
Because Kiwi contains protease enzymes which break
up proteins they get destroyed. It is also why it can hurt
if you eat too many - they enzymes start to
breakdown the proteins that make up your mouth.
The salt makes the DNA stick together. The DNA
doesn't dissolve in alcohol so it coagulates to form
a jelly like substance which you can pick up.
Use the website provided below to study the simple extraction process.
http://www.daveansell.co.uk/?q=node/31
Experiment: Fruitful DNA Extraction
Aim:
To extract DNA from kiwifruit
Introduction:
DNA is contained inside the nucleus of a cell. To extract the DNA, the cells have to be heated
to high temperatures. The steps below explain how this is achieved.
Background Information:
DNA is the genetic information in the organisms. We can extract DNA from the tissue using a
very simple procedure.
Equipment:
A kiwi fruit
5g (5 mls) washing up liquid or hand soap 2g (1/2 tsp) salt
100ml tap water
100ml of ice cold alcohol (methylated spirits work well). Put in freezer for at least 30 mins.
Kettle
3 jars - 100ml and 2 x 150ml beakers
Large basin – 250ml beaker
Bowl (or mortar and pestle) to mash the kiwi with sieve or coffee filter paper
Knife (be careful!)
Method:
1 - Peel the kiwi fruit and chop it into small chunks. You don't want the skin because it's
mostly dead and doesn't have much DNA in it.
2 - Put the chunks in a bowl (mortar) and mash the kiwi as much as you can. This is to
break up some of the cells and provide a large surface area over which to extract the
DNA
.
3 – In the 150ml beaker mix together the washing up liquid, the salt and the tap water
and stir slowly until the salt has dissolved. Don't stir too fast or else you'll get lots of
bubbles! This mixture is also known as an extraction buffer.
4 - Add the extraction buffer to the mashed-up kiwi in the mortar and MASH! The more
you mash, the more DNA you will get out at the end. Pour into 100 ml beaker.
5 - Incubate the kiwi and buffer mixture (in 100ml beaker) at 60 degrees Celsius for 15
minutes in the 250ml beaker (incubator/water bath).
To make your own incubator/water bath, take a large basin (250ml beaker) and 1/4 fill
it with boiling water from a kettle. To reduce the temperature, add about the same
amount again of normal tap water. Using a thermometer will help you reach a more
precise temperature. Carefully put the jar (100 ml beaker) with the kiwi into the
incubator (250ml beaker) and leave to stand for 15 minutes. Incubation helps to break
up the cells further and starts to degrade some of the cell's proteins.
6 - Remove the jar (100ml beaker) from the incubator and filter the kiwi mixture
through a fine sieve or coffee filter paper into another jar/beaker. This removes all
the unwanted lumps and bits of kiwi fruit. You should be left with a green liquid, and
this contains the kiwi fruit DNA.
7 - Take the ice-cold alcohol and pour it slowly down the side of the jar/beaker. The
alcohol will form a transparent layer on top of the kiwi mixture, as the alcohol is less
dense.
Where the layer of ice-cold alcohol meets the kiwi mixture underneath, you will see a
white jelly- like substance forming. You can hook the DNA out with a hook made from
a paperclip or something similar.
All living things are made up of cells, and each cell contains a complete copy of the organism's DNA.
So you could extract DNA from any kind of living material.
http://www.thenakedscientists.com/HTML/content/kitchenscience/exp/how-to-extract-dna-from-a-kiwi-fruit/
