Sequencing PCR products - (Jan/22/2001 )

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 ( @ 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-

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

-jadefalcon-

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