I am trying to amplify amylase gene from Bacillus licheniformis but the problem is reproducibility of the desired band.
Could anyone guide me?
You may have to look the steps and the types of buffers included in the kit. But in general, the viral DNA kit can’t be used for bacterial DNA since both the organisms don’t share the overall structure that surrounds genome. Bacterial walls are too complex and needs harsh treatments to get burst.
Are you saying that you tried this viral kit (for bacterial DNA) and now the PCR is not reproducible? If so, no wonders…! Instead of using the viral kit, go for proper method or suitable kit. You can do it manually as well if the kit is not available. I mean you should not expect it to be reproducible while you have purified the DNA with such tricks.
Lastly, you may have to elaborate your query like size of the gene you tried to amplify? What variations have you already tried? What wrong are you getting (no bands, smaller band or very thin band)..? then only you will get perfect suggestions.
Good Luck
Thank you For your kind responses,
yes, I was trying to amplify the gene of interest using vial DNA extraction kit. I got the band of interest (1.5 KB) with adequate thickness.... but when repeated, could'nt get that again.... Now I will use the bacterial DNA extraction kit ..hope it will work...
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