2005-2006 AP Biology Biotech Tools Review 2005
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Transcript 2005-2006 AP Biology Biotech Tools Review 2005
Biotech Tools Review
AP Biology
2005-2006
Biotech Tools Review
Recombinant DNA / Cloning gene
restriction enzyme, plasmids, bacteria
DNA library
cDNA library
DNA hybridization
Gel electrophoresis
RFLP
Southern blotting
PCR
Sanger sequencing
Microarray
AP Biology
2005-2006
You want to produce human insulin
cheaply for diabetics
Find your gene
collect mRNA from pancreas cells
What to do with the mRNA?
AP Biology
2005-2006
You want to produce human insulin
cheaply for diabetics
What to do with the mRNA?
make DNA
mRNA cDNA
reverse transcriptase
insert cDNA into bacteria using plasmids
restriction enzyme
ligase
transformation
How do you find which bacteria carry
the recombinant plasmids?
AP Biology
2005-2006
You want to produce human insulin
cheaply for diabetics
How do you find which bacteria carry
the recombinant plasmid?
amp resistance gene on plasmid
LacZ gene plasmid
EcoRI
BamHI
HindIII
LacZ gene
plasmid
amp
resistance
AP Biology
2005-2006
You want to produce human insulin
cheaply for diabetics
How do you find which bacteria carry
the insulin gene on the plasmid?
make an insulin gene probe
tagged probe
hybridization
labeled probe
genomic DNA
3’
AP Biology
G A T C AG T A G
CTAGTCAT C
5’
2005-2006
What if you want to find out the full
sequence of the human insulin gene?
Sanger sequencing
AP Biology
dideoxynucleotides
2005-2006
What if you want to make a lot of
copies of your sequenced gene?
PCR
AP Biology
thermocycler
primers
Taq polymerase
2005-2006
You want to see if 2 people have
differences in their insulin genes?
Gel electrophoresis & RFLP
restriction digest
run samples on gel
see if there is a polymorphism
AP Biology
2005-2006
What if I want to locate a sequence
on an electrophoresis gel?
Southern blot
AP Biology
tagged probe
2005-2006
You want to test effect of drug on
gene expression in pancreas?
Microarray
collect mRNA
mRNA cDNA
hybridize on GeneChip
AP Biology
2005-2006
Any Questions??
AP Biology
2005-2006