Kindly explain the principle that guide the triming of e. coli 16S rRNA gene sequence while trying to customize for the region we use. I have gone through your explanations on how to customise for using V3 rather than V4 in the mothur SOP. My problem is how to identify where to insert the forward primer and where to put the complimentary sequence of the reverse primer.
Hi - Do you understand how PCR works and how the primers anneal to the sequence? You do the same thing, but on the computer with your primer sequences and the E. coli sequence.
pat
Dear Pat,
THank you so much for your support, I have done that and worked well.
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