Hi everyone. I´m having problems to amplify the GyrB gene in bacteria. I did the DNA extraction, and I could amplify the 16s without problems, but when I try with GyrB I can´t get any product. I´m using the UP1 and UP2 primers with 60° of annealing temperature. I tried different quantities of DNA, but I think that is not the problem, because with 16s it works very well. I have tried adding BSA and nothing happens.
Do you have some idea what is the problem ??. If somebody can help me..
Thanks a lot !!
Hi walter, I used the UPs primers, they were designed as universals. The first article was presented by Yamamoto & Harayama, 1995 in a taxonomic study of Pseudomonas strains. But, I think now they are widespread used primers. I dont think the problems could be the primer sequence. I have tried both, degenerate and non degenerate sequence without results. I thought first in the DNA quality but, it works with 16s, so should be works now too.
Whatever. thanks a lot for your comments, I will be trying to do that.
Greetings..
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