Gel electrophoresis

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suzy06
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Gel electrophoresis

Post by suzy06 » Mon Aug 13, 2007 12:42 am

Hi everyone, I have a very quick question which is worth alot of marks ... I would be very thankful if someone helps me with it ... and please answer soon !!

question: after running on a gel, one finds that there is no insert and there is only one band on th gel and it has the same size as the vector. why this hapened ? explains ... and also comes to know that he amplified the false positive colony ....

SororSaudade
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Post by SororSaudade » Mon Aug 13, 2007 4:24 pm

it seems your primers also anneal in the vector alone... you can try to increase the Ta.

I supose you did like a negative control of the PCR (so that you can discard the contamination hypothesis).

suzy06
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Post by suzy06 » Mon Aug 13, 2007 4:43 pm

hey, can you explain a little bit more about the negative control of PCR ?

SororSaudade
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Post by SororSaudade » Mon Aug 13, 2007 5:05 pm

Is just a control without DNA (or with a sequence that you absolutely know that does not amplify eith the primers you're using). It allows you to know if your mix is contaminated (it is if you get any amplification in this sample)

Any other question, just ask :)

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