DMSO Contamination in PCR - (Jun/09/2006 )
Hi,
I am trying to see the effect of DMSO on PCR reactions and fragment analysis.I use the FTA cards from whatman and spot a certain amount of cells onto FTA paper and punch out a disk from it and set up PCRs.Does the DMSO in the freezing media can be carried in to PCRs after 3 washes with FTA purification reagent and 1 was with TE?
Also does anybody know what kind of effect can DMSO have on fragment analysis,suppose if I use the PCR product which has DMSO.
Any suggestions will be greatly appreciated.
Thanks,
Megha.
Hi,
I have done PCR using DMSO as one of the constituents. It is used when the template is highly GC-rich. And as far as I know DMSO does not cause any problem in amplification. Usually you would see that on increasing DMSO % in the reaction the amplified product would increase and then decrease; sort of a bell-shaped curve.
Sunil
I have done PCR using DMSO as one of the constituents. It is used when the template is highly GC-rich. And as far as I know DMSO does not cause any problem in amplification. Usually you would see that on increasing DMSO % in the reaction the amplified product would increase and then decrease; sort of a bell-shaped curve.
Sunil
Hi Sunil,
Let me rephrase my question does DMSO inhibit the fluorescent allele detection with the ABI PRISM 310?
Will it interfear with the dye chemistry or intensity of peaks.....
I feel since freezing media has DMSO it can be carried into PCR and flouresent allele detection even after several washes with FTA reagent. My primers are not GC rich.I am not worried about increasing my PCR yield I am only concerned about it after PCR when I run PCR products in ABI 310.
I would appreciate your reply,
Megha.
I have used DMSO in my sequencing reaction PCR (for shRNA) and they didnt disturb the signal. U get good peaks if the DMSO is added for certain sequences.