From 3-21-15.
Attempted to amplify the 16S rRNA gene of the five P. larvae honey isolates and the roach trachea isolate using the 8F and 16Srev (from IDT) primers using a thermocycler parameter I had previously developed based off a paper that used the 8F primer to amply the region. HOWEVER, the gel was completely blank, except for the DNA marker, indicating the amplification was not a success.
I have not been able to successfully amply the 16S using these primers and those parameters in the past, but I believe the problem to be the template DNA. I am confident that the DNA I used as a template is adequate for amplification as it was used successfully previously using the AFB primers. I am now convinced that the issue is either my amalgamated use of primers or the thermocycler parameters. I will attempt to remedy in the near future as I would like to send the 16S for sequencing soon.
//EWW
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