Wrong PCR band size

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Wrong band size

Im having problem with a slightly wrong band size for one of my pcr products. I newly extracted the DNA therefore what could possibly be other reason besides template contamination?

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Rajib Roychowdhury

Please check the pH of storage buffer or do PCR with DNA that dissolved in MilliQ water.

Prashant Bhaskar Kale

whether it is higher than expectation or lower?

if higher/multiples then, reduce MgCl2/Primer/template con.

Asdren Zajmi

the band is slightly lower by 10-20bp. I use 100bp dna ladder.

It’s ok till the difference is of 10-15bp --->you carry on to next protocol...

The reason: this may be due to software handling (adding markers to gel picture….) and run parameters like gel/ladder used.

good day...

thank you!

Hi BH Kong. I haven't sent the samples for sequencing yet but will be sending this week. Thank you !

Hi BH Kong, i don’t think so!!!!, Please explain your case; What was the difference between observed and expected size?

hi Asdren Zajmi, if you got your samples sequences --->Just BLAST them on NCBI