Band in CNT

I recently ran a cycle course PCR reaction on 09/15 using the fancy thermocycler on the first bench. a strange band appeared at about 100bp in my NTC lane when i ran the gel of my amplified samples. The picture is a bit blurry:the lanes on the top left are for wash 3 (cycles 9, 13, 15, and 20). the lanes on the top right are for wash 0 (cycles 9, 13, 15, and 20). the lane on the bottom far left is the NTC and the lanes on the bottom right are for elution 1 (9, 13, 15, and 20). I am well aware that not much appears to be amplified here, which is why i am running another cycle course on my wash 0 and another NTC to see if the band appears again, as its original source is not apparent from the gel.

when running the gel, i used a rig that already had the TBE in it from someone else's previous gel run (no clue who) and im pretty sure it was one of the orange rigs -- although i doubt that matters at all.

From the best of my powers of deductive reasoning, the only specific information on aliquots that i could say for sure is:
-4mM dNTP (2/8/10)
-diH2O (9/7/11 ABR)
-100bp ladder (9/2/11)
-6X orange dye + EtBr (10/27/09 SKP)
none of the 10xPCR buffer or primer aliquots are labeled with dates.

All in all, i won' be sure if this is an actual artifact like Gwen was talking about until i complete my wash 0 and NTC cycle course check, which should be tomorrow sometime.
-Cori Booker

1 comment:

Gwen Stovall said...

Hi Cori,
Thank you very much for posting these results and helping us understand more about this "amplification artifact" that appears in PCR reactions when template is NOT added. Which primers did you use?

Thank you,