Hi eveyone, i have a few very basic questions that are buging me: - The emulsion PCR performed to create a template results in amplicons attached to a bed, which in turn goes to the pico-well and sequencing is performed afterwards. Such amplicons habe a forward strand attached to the bead and a reverse strand "free" that detaches after denaturation, leaving only the forward (5-3) starnd as the sequencing template, with he P1 adapter at 5' and the A adapter at 3'. Correct?
Sequencing will ocurr after a primer anneals to the 3' end of the template. That region corresponds to the A adapter and the sequence occurs towards the bead. As a result, bases are called from the primer annealed to the A adapter and will finish in the last nucleotide of the P1 adapter (on the bead). Is this correct?
Adapter trimming therefore should be performed at the 5' end (A adapter) and at the 3' end (P1 adapter)?
Why is always talked about trimming only the 3' end?
I know is quite a long post, however i am new at NGS.
Thanks in advance, Sergio Sergio.pv is offline