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| Hello, 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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| hygromycin , primers |
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| Thread | Thread Starter | Forum | Replies | Last Post |
| hygromycin primers | SVemuri12@gmail.com | Protocols and Methods Forum | 2 | 10-11-2007 11:06 AM |
| hygromycin primers | Vince Mulholland | Protocols and Methods Forum | 0 | 10-09-2007 01:12 PM |
| hygromycin primers | Analia Alet | Protocols and Methods Forum | 0 | 10-08-2007 09:24 PM |
| [Arabidopsis] GUS and Hygromycin resistance gene primers | Michael Sullivan | Arabidopsis and Plant Biology | 0 | 03-21-2007 06:34 PM |
| GUS and Hygromycin resistance gene primers | senani dayabandara | Arabidopsis and Plant Biology | 0 | 03-21-2007 05:24 AM |