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Biotechnology Unit

Biotechnology involves manipulating existing genetic systems in order to achieve a desired outcome.

http://www.dnai.org/

3.5 Genetic Modification and Biotechnology

things to remember from class 1:

- PCR, or amplifying DNA, is required for all other DNA technologies

Notes from Bioninja

Understandings

- PCR can be used to amplify small amounts of DNA


o PCR stands for polymerase chain reaction
o Its an artificial method of replicating DNA under laboratory conditions
It amplifies a small sample of a specific sequence of DNA into large quantities
Each reaction cycle DOUBLES the amount of DNA
A standard PCR sequence of 30 cycles creates over 1 billion copies, or 2^30
copies.
o Stages of PCR
Denaturation
DNA sample is heated to separate it into two single strands
This is at 95 degrees Celsius for 1 minute
Annealing
DNA primers attach to the 3 ends of the target sequence
This is at 55 degrees Celsius for 1 minute
Elongation
A DNA polymerase that can stand heat (its called Taq) binds to the
primer and copies the strand
This is at 72 degrees Celsius for 2 minutes
o Then the large quantities of DNA are isolated and manipulated using different
techniques.
- Gel electrophoresis is used to separate proteins or fragments of DNA according to size
o This is the isolation part
o So the DNA samples are put into a block of gel and then theyre electrocuted (an electric
current goes through them)
o That makes the samples move through the gel, right?
o Right. And then smaller samples move quicker through the gel, cause they can
o So what this does is it makes the samples separate, cause theyre moving at different
speeds
o Oh and it goes from the negative terminus, or the cathode, to the positive terminus, or
the anode.
o But DNA and protein separation follow different protocols
DNA separation
So the DNA could be cut into fragments with something called
restriction endonuclease, and obviously different DNA samples make
different fragment lengths cause theyre all different sizes
So once these fragments are made they separate because DNA is
negatively charged, right?
o Right. Cause DNA has a PO4 3- on each nucleotide, and that
phosphate group is negative.
Okay, so then DNA samples are put into a gel called agarose gel
Then this funky thing called a complementary radiolabelled
hybridization probe, identifies a specific sequence
Then it transfers it into a membrane
- DNA profiling involves comparison of DNA
- Genetic modification is carried out by gene transfer between species
- Clones are groups of genetically identical organisms derived from a single original parent cell
- Many plant species and some animal species have natural methods of cloning
- Animals can be cloned at the embryo stage by breaking up the embryo into more than one
group of cells
- Methods have been developed for cloning adult animals using differentiated cells