Does anyone know how to overcome internal blocks in PCR product templates used for sequencing. I've heard that betaine or DMSO can be used, any comments?
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thegradstudent
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Posted 27 March 2004 - 03:59 PM
| QUOTE ( @ Jan 22 2001, 10:00 PM) | |
| Does anyone know how to overcome internal blocks in PCR product templates used for sequencing. I've heard that betaine or DMSO can be used, any comments? [/quote] I have heard that alcohol could be an importantfactor in this,so you have to be sure to eliminate before eluting your nucleic acid. Also adding DMSO or betaine can help
TheGradStudent
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jadefalcon
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Hi - simple advice:
try again... we sometimes have to sequence past a "G"-run...2 out of 3 identical reactions will break off at this point, our sequencer says: "No Way we can get over that!", and we try it again, and it works - same conditions, reagents, etc. sometimes you just have to be lucky! as thegradstudent wrote above ethanol will inhibit your sequencing reaction, so be sure to get a clean template for your sequencing! Mike Edited by jadefalcon, 28 March 2004 - 10:07 AM.
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