Cell Bio Exam 3 Dimario, chapter 6 Exam 100%
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Terms in this set (130)
PCR amplifies specific DNA exponentially
What is Polymerase Chain
Reaction?
- most powerful
Forward and Reverse primers select the specific
How does PCR work? sequence of DNA to be amplified (Primer is an
oligonucleotide)
in vitro organic synthesis a 5' OH
is backwards using
In PCR denaturation 95
occurs at ?
In PCR annealing of 55
primers occurs at?
Elongation of primers 72
occurs at? special heat
stable dna polymerase
first cycle in PCR depends Tm of the primers
on?
How is the primer able to Primers are in vast molar excess compared to the
attach to the DNA without original template DNA (they serve as starting points);
the single strands this allows for multiple cycles
reannealing to each
other?
, Why would you design allows you to ligate with to a plasmid vector with the
your primers to include same restriction sites.
restriction enzyme sites?
What is directional ligate with plasmid with sticky ends after cut with
ligation? restriction enzymes
What is A Newer Method gipson assembly
for Producing
Recombinant Plasmids:
several PCR products all at once in the correct
gipson assembly ligates
orientation
compatible sticky ends
Gipson assembly anneals
- 3 extension with a polymerase to fill in gaps
What is Maxam-Gilbert end labeling DNA fragments, then chemically
Sequencing? cleaving the DNA
Maxam and gilbert dna 32P added to 5' ends by a kinase
sequencing technique
uses what end labeling? - Polyacrylamide gel/autoradiography
and what material?
Sanger sequencing uses? dideoxy-ribonucleosides triphosphates (ddNTPs)
and sequences how - up to 500-600 bases per run
much?
No 3' hydroxyl means? no further synthesis (gel termination)
Solved
Save
Terms in this set (130)
PCR amplifies specific DNA exponentially
What is Polymerase Chain
Reaction?
- most powerful
Forward and Reverse primers select the specific
How does PCR work? sequence of DNA to be amplified (Primer is an
oligonucleotide)
in vitro organic synthesis a 5' OH
is backwards using
In PCR denaturation 95
occurs at ?
In PCR annealing of 55
primers occurs at?
Elongation of primers 72
occurs at? special heat
stable dna polymerase
first cycle in PCR depends Tm of the primers
on?
How is the primer able to Primers are in vast molar excess compared to the
attach to the DNA without original template DNA (they serve as starting points);
the single strands this allows for multiple cycles
reannealing to each
other?
, Why would you design allows you to ligate with to a plasmid vector with the
your primers to include same restriction sites.
restriction enzyme sites?
What is directional ligate with plasmid with sticky ends after cut with
ligation? restriction enzymes
What is A Newer Method gipson assembly
for Producing
Recombinant Plasmids:
several PCR products all at once in the correct
gipson assembly ligates
orientation
compatible sticky ends
Gipson assembly anneals
- 3 extension with a polymerase to fill in gaps
What is Maxam-Gilbert end labeling DNA fragments, then chemically
Sequencing? cleaving the DNA
Maxam and gilbert dna 32P added to 5' ends by a kinase
sequencing technique
uses what end labeling? - Polyacrylamide gel/autoradiography
and what material?
Sanger sequencing uses? dideoxy-ribonucleosides triphosphates (ddNTPs)
and sequences how - up to 500-600 bases per run
much?
No 3' hydroxyl means? no further synthesis (gel termination)
