PCR and primer dimers...

[darkwng]

I have primer dimers and not much product.... what should I do to fix this?

Darkwng

[MicroKiller]

Try using less primers for ur pcr.. its possible u have excess primers in ur PCR reaction. wht is the amount of sample and primer u are using.. You may need to use more sample for your PCR. Question:

1. Is your primers specific to your gene of interest.?
You could try running an in-silico pcr to determine if your primers are suitable.

[PCR-Man]

Hi Darkwng,
how did u design the primers? how much dna and how much primer do you put in? what is the annealing temp and time u use?

[darkwng]

Hi PCR MAN!
thank you for help!

I put in 100uM of primer at annealing temp 60C for 1 minute

thank you

[saly]

please if possible , send to me to also, solve my problem.
Please i need the help in my problem:
i design primer but when i made PCR, i found may be formation to
primer dimer under the louding dye but the band of primer dimer under
the negative control ( without RNA ) was small in size than the band
under the sample ( more thick and may be 2 band more closed ). i
belived fond 2 band in sample because the negative control and sample
contain same band in the below but found band upper it in the sample
( may be more thickness )at the same conditions. i tried at 47, 52,
55oC annealing. all tempreature give same result with diffrence in 47
the band under negative control more faint but at 55 the band under
sample more thick and light.
please, i need to know this difference meanse any thing or the
difference in size between the band under control and sample don't
means any thing and the 2 bands are primer dimer only.
Thanks for you

[saly]

please can i find any body reply to my question.