|well good news its working now. i don't know how but i get the feeling it was a contamination prob.as u said i kept the tips and pipettes under UV. but still i get a bit of a smear after the band. but its very light. |
i have to send these for sequencing so i'm just wondering whether even if i have a bit of a smear whether it ll b a prob during sequencing..
primers ..well its working again so can't b primers..but anyway we are going to order new primers. so have to c what effect will come with the new primers.
my usula MgCL2 conc in 2mM and today i lowered it to 1.5mM but the smears hasn't changed..can't figure out what it is..i'm doing a HOt start PCR also..
hey that's good. but try to figure
out that what went wrong. my problem is that i'm getting the band in the positive control but not in the samples. i don't thnk that are degraded because these are extracted very recently.can anyb'dy have some idea about purification
of these DNA
samples. i also got the band of just the DNA
on .7% gel
but what is gng wrong i don't know. i changed my DXTPS but not worked as i thght.but smearing is less in positive control. i diluted my samle also but didn't worked either. i thnk there is possibly some reagent
during dna extraction
which is affecting the whole pcr
. give me some idea.