SEQanswers - View Single Post - 'Assembly PCR' for homemade amplicon library prep

DNA_Dan · 11-13-2014, 02:37 PM

I think Illumina recommends a two-step approach. Fusion primers with barcode and sequencing primer locations, then a second PCR with P5/P7 ends being added. If you do it all at once it may get ugly. Also if you are low diversity adding in a N+1, N+2 linker in there greatly helps diversity.

11-13-2014, 02:37 PM	#2
DNA_Dan Member Location: Montana Join Date: Nov 2008 Posts: 21	I think Illumina recommends a two-step approach. Fusion primers with barcode and sequencing primer locations, then a second PCR with P5/P7 ends being added. If you do it all at once it may get ugly. Also if you are low diversity adding in a N+1, N+2 linker in there greatly helps diversity.