Download AP-ppt-PCR

Problems
Not all bacteria pick up
plasmid-how do we
distinguish?
Annealing of human
DNA to plasmid is
random-how do we
distinguish which
plasmids have human
DNA?
How do we find the
gene
we want in a library?
Probing!
It’s like fishing For
DNA
Relies on
complementarity of
The double strand
Sometimes there just isn’t enough
Of the DNA we want to work with!
The answer is PCR
Polymerase Chain Reaction
http://dnalc.bii.a-star.edu.sg/shockwave/pcranwhole.htm
l
DNA
sequencing
http://www.pbs.org/wgbh/n
ova/genome/sequ_flash.ht
ml
Southern Blotting-used to identify
genes/sequences
http://dnalc.bii.a-star.edu.sg/shockwave/southan.htm
Copy DNA(cDNA)
DNA microarray analysis
Microscopic DNA
attached to solid surface
like glass, silicon
Allows us to answer
questions about gene
activity



Different stages of
development
Different tissues
Healthy vs diseased
tissues
RFLP’s-Restriction Fragment Length Polymorphisms
Variations in the length of fragments resulting from action
by a specific restriction enzyme
uses
transposons
“Jumping genes”
Approx. 45% of human genome
Can cause mutations
Can be used to trace genetic origin
SAFETY, SAFETY, SAFETY!!!
THINK!
No food or drink anywhere!!!!
Don’t touch face, doorknobs, goggles,
books, faucet handles, etc with gloves on
Clamshell technique
Wipe surfaces thoroughly w/ ethanol spray
So what about our lab?
We artificially transformed
bacteria
Gene comes from jelly fish!
The Arabinose operon
How do we know which
bacteria took up plasmid?
What is required in the agar
to make them “glow”?