Biology Forum › Molecular Biology › Gel electrophoresis
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- August 13, 2007 at 12:42 am #8079
suzy06ParticipantHi 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 ….
- August 13, 2007 at 4:24 pm #75161SororSaudadeParticipant
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).
- August 13, 2007 at 4:43 pm #75162suzy06Participant
hey, can you explain a little bit more about the negative control of PCR ?
- August 13, 2007 at 5:05 pm #75164SororSaudadeParticipant
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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