Hi, If the PCR used to work but is not working anymore, you may have some PCR inhibitors in your samples. Diluting them in water, about 100 times, can help (but trying different dilution factors, e.g. 10, 100 and 500 may be a good idea). Alternatively, your template concentration may be too low or your DNA may be degraded.
If this is a new PCR, you may lower the annealing temperature or increase the concentration of primers (generally in the 0.2 µM - 0.8 µM range for the final concentration). However, in the first case, you may loose specificity and in the second, you may generate primer dimers. I hope this helps. Sébastien On Tue, Apr 6, 2010 at 9:05 AM, Christian Praetorius <[email protected]> wrote: > LeboMad <[email protected]> wrote: > > >have done all troubleshooting there is out there I can't seem to get > >it right. I normally get nice clear bands but for some reason I get > >very faint bands if any at all. Can anyone suggest anything. I've got > >a student that needs some PCR runs and needs result as in yesterday. > > The nyou should describing *exactly* what your procedure is and what > you tried to troubleshoot. > > Christian > > -- > X-no-Sig: yes > _______________________________________________ > Methods mailing list > [email protected] > http://www.bio.net/biomail/listinfo/methods > -- Sébastien Vigneau Lab: Marisa Bartolomei's laboratory - 360 Clinical Research Building - 415 Curie Blvd- Philadelphia - PA 19104-6148 - USA -- phone: +1-215-898-9277 -- fax: +1-215-573-6434 _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
