Techniques Flashcards by emily chetwin | Brainscape

Q

What happens if primer annealing temp is wrong?

A

Too high - primers melt off

Too low - off target amplification

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Q

Reagents ended for PCR

A

template DNA
primers
dNTPs
mgCl2
standard buffer
taq
water

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Q

Formula for melting temperature (Tm)

A

Tm = 4(G+C) + 2(A+T) C

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Q

What is the general annealing temp for a primer?

A

5 degrees lower than the Tm

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Q

At what rate does taq extend?

A

100 bases/sec

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Q

Stages of PCR

A

Denaturation
Amplification
Extension

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Q

Disadvantages of Sanger sequencing

A

not high throughput

- high cost per base

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