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hygromycin primers

hygromycin primers - Arabidopsis and Plant Biology

hygromycin primers - Discuss Arabidopsis and Plant Biology Research.

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Old 10-07-2007, 10:11 PM
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Default hygromycin primers


Those those who have used hygromycin primers:

I am trying to amplify plant genomic DNA (WT and mutant lines) using
Hyg primers. However, so far, I have had no luck. My DNA template is
ok because I saw DNA pellet. I think it is the annealing temperature
on which I am doing the PCR. My question is, what annealing
temperature is ideal for these primers to hybridize? I have been doing
PCR reactions with annealing temperatures ranging from 58-61 degrees
Celsius, with no luck. So far, I've been observing different annealing
temperatures, but are there suggestions to other things I should look
at to make these primers hybridize to the DNA? The primer sequences
are below.

Forward primer:
5' AGC TGC GCC GAT GGT TTC TAC AA 3' Tm = 61.70 deg Cel

Reverse primer:
5' ATC GCC TCG CTC CAG TCA ATG 3' Tm = 58.84 deg Cel

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