test 3 prep Flashcards
what contributes to the high fidelity of DNA
DNA repair enzymes, base pairing rules, and proofreading capabilities of DNA polymerase III
what does DNA polymerase require to start synthesizing a new strand?
free 3’ end. free template strand. dNTP. Mg++
how can you tell if a baby belongs to a parent by using VNTR?
each VNTR fragment of the child must match one from the parents
what is the relationship between the base sequence of coding strand and base sequence of mRNA?
they are the same, except the thymines are replaced with U’s in RNA
determine size of mRNA- use what technique?
northern blotting
identify a colony from a cDNA library using an antibody?
gene expression screening
identify bacterial colonies who have taken up plasmids via transformation
antibiotic resistance analysis
expression screening
detect protein of interest using antibodies. sticks to tubulin.
Screening via hybridization:
use radioactive probe to identify specific sequences
DNA moves towards (+/-) end
+ end. it’s negatively charged
Southern Blotting
use hybridization to see which fragment in electrophoresis was of interest; identify DNA fragments complimentary to the probe
what do you do if you want to find the size of RNA
northern blotting
antibiotic resistance analysis is used for what process
identifying bacteria colonies that have taken up plasmids via transformation.
in PCR, which end do primers face
3’
steps of PCR
denature, anneal, repeat/extend
Chain termination technique for DNA sequencing
a method of DNA sequencing based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during DNA replication. stops chain, and then you can see where it stopped. how they mapped human genome
what is the blue white test and what is it used for?
screens for plasmids with inserts (inserted DNA sequences)
how would you produce DNA versions of mRNA sequences
cDNA synthesis
discriminate between different allelic forms of B-globin?
RFLP
in blue/white test- what exactly is turning blue? what gene is on/off?
lac Z gene . active lac z=blue, inactive (has plasmid) turns white
relationship cAMP to CAP and what does it do
cAMP binds to CAP in absence of glucose, changing its conformation, so that CAP can bind to activator site
which enzyme turns indicator dye blue?
B-galactoside