Good day everyone. I've carried out mPCR for the confirmation of Non Typhoidal Salmonella using DNA extracted kit( G-spin from INTRON ) .While all the 5 strains (S. Typhimurium, Enteritidis, Heidelberg, Agona, welteverden) gave the expected bands when ( Lane 4-8), the postive control, the pool DNA (lane 3)was missing two of the expected band( S.Agona and Heidelberg). I was wondering what could be the possible cause as the DNA used for the pool DNA are of the same sample used in the single lanes. All concentration of the DNA are standardized and the reagents are from the same master mix. I will attach the gel pic along with the questions. Thank you in advance for the replies. I really appreciate it.
lane 1: ladder
Lane 2 negative control
Lane 3 positive control (pool DNA)
Lane 4-8 samples ( S. Typhimurium, Enteritidis, Heidelberg, Agona, welteverden )
hi freddy, from your question i understood that lanes 4-8 samples were pooled and ran in lane 3. is it true?
can you please specify the source of sample, is it culture are from human samples?
if you kindly reply to these questions, then we can think of some interpretations. because we encountered similar type of problem in our lab
Hi Rishi. Yes the lane 3 was pooled from Samples 4-8.
As for your next question , the samples are from ATCC.
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