B-galactosidase
lactose
galactose
glucose - ANSWER LacZ codes for ________ which hydrolyzes _______ into
______ and_____
chaotropic - ANSWER small molecules used to denature proteins (urea,
guanidinium chloride); bind to water and reduce hydrophobic effect
The melting temperature formula will be given if we need it, but how do you
calculate the annealing temperature if you have the melting temperature -
ANSWER subtract between 2-5 degrees from the melting temperature
Taq Polymerase can add _______bp/min - ANSWER 1000
_________ is a cofactor for Taq - ANSWER magnesium
What replaces helicase in PCR? - ANSWER Heat
primers bind to the _____ end of DNA strands - ANSWER 3' end
Longer primers require ________ melting temperatures which can prevent..... -
ANSWER higher
mispriming
In order to do PCR you need _____,_____, and _____ - ANSWER dNTP
Primers
Polymerase
Which nucleotide bond is stronger and why? - ANSWER G-C because it forms
3 H-bonds whereas A-T only forms 2 H-bonds
How long is the LacZ gene? - ANSWER ~3000bp
, Optimal prier is around ____bp and has a melting point between ____-
____degrees celsius - ANSWER 20
55-65
Total cation concentration for calculating Tm is usually _____ - ANSWER
50mM
How do you prevent a plasmid from reattaching to itself after it has been cut
using restriction enzymes? - ANSWER You dephosphorylate the sticky ends
of the cut plasmid using alkaline phosphatase.
This involves removing the terminal phosphate groups of the DNA so that no
phosphate is available.
If you increase the % agarose of a gel tray then the DNA will migrate ______ -
ANSWER slower
What is in a TBE buffer? - ANSWER - EDTA - which chelates divalent
cations (specifically Mg2+). Magnesium is often required by enzymes that
break up DNA, so EDTA protects DNA from degredation.
- Tris basic salt - provides conductivity
- Boric Acid - Lowers pH so DNA does not hydrolyze and retains correct
negative charge on DNA
DNA migration speed is proportional to...... - ANSWER 1/log(#of bp)
Gel separation of DNA is most effective when DNA is between ____bp and
____bp - ANSWER 10 and 50000 (50kb)
DNA migrates towards the _________ pole of the eletrophoresis chamber
because it is _________ charged. - ANSWER + (anode)
negatively
DNA is _______ charged and silica is ____ charged. DNA attaches to silica via
________ which is found in the __________ - ANSWER negatively
negatively
guanidium HCl
Binding buffer
lactose
galactose
glucose - ANSWER LacZ codes for ________ which hydrolyzes _______ into
______ and_____
chaotropic - ANSWER small molecules used to denature proteins (urea,
guanidinium chloride); bind to water and reduce hydrophobic effect
The melting temperature formula will be given if we need it, but how do you
calculate the annealing temperature if you have the melting temperature -
ANSWER subtract between 2-5 degrees from the melting temperature
Taq Polymerase can add _______bp/min - ANSWER 1000
_________ is a cofactor for Taq - ANSWER magnesium
What replaces helicase in PCR? - ANSWER Heat
primers bind to the _____ end of DNA strands - ANSWER 3' end
Longer primers require ________ melting temperatures which can prevent..... -
ANSWER higher
mispriming
In order to do PCR you need _____,_____, and _____ - ANSWER dNTP
Primers
Polymerase
Which nucleotide bond is stronger and why? - ANSWER G-C because it forms
3 H-bonds whereas A-T only forms 2 H-bonds
How long is the LacZ gene? - ANSWER ~3000bp
, Optimal prier is around ____bp and has a melting point between ____-
____degrees celsius - ANSWER 20
55-65
Total cation concentration for calculating Tm is usually _____ - ANSWER
50mM
How do you prevent a plasmid from reattaching to itself after it has been cut
using restriction enzymes? - ANSWER You dephosphorylate the sticky ends
of the cut plasmid using alkaline phosphatase.
This involves removing the terminal phosphate groups of the DNA so that no
phosphate is available.
If you increase the % agarose of a gel tray then the DNA will migrate ______ -
ANSWER slower
What is in a TBE buffer? - ANSWER - EDTA - which chelates divalent
cations (specifically Mg2+). Magnesium is often required by enzymes that
break up DNA, so EDTA protects DNA from degredation.
- Tris basic salt - provides conductivity
- Boric Acid - Lowers pH so DNA does not hydrolyze and retains correct
negative charge on DNA
DNA migration speed is proportional to...... - ANSWER 1/log(#of bp)
Gel separation of DNA is most effective when DNA is between ____bp and
____bp - ANSWER 10 and 50000 (50kb)
DNA migrates towards the _________ pole of the eletrophoresis chamber
because it is _________ charged. - ANSWER + (anode)
negatively
DNA is _______ charged and silica is ____ charged. DNA attaches to silica via
________ which is found in the __________ - ANSWER negatively
negatively
guanidium HCl
Binding buffer
