Nanopore library on top of an Illumina library

drops · 11-30-2020, 09:33 AM

Hi all,
we don't have the original aliquote of an old sample, but we have an Illumina Nextera Flex library in the freezer.

I am aware that the ends are y-shaped, correct? Anyone of you have a fancy protocol to remodel/fill/modify those ends that I can do a Nanopore library on top?

Thx!

luc · 11-30-2020, 09:45 AM

No remodeling needed.
All Nextera libraries are PCR amplified to be sequenceable. After the PCR amplification they are wonderfully double-stranded throughout and should be blunt-ended (if a high-fidelity polymerase was used).

BTW, Truseq-style libraries have Y-adapters; Nextera/transposase libraries work differently.

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11-30-2020, 09:33 AM	#1
drops Junior Member Location: Barcelona Join Date: Nov 2011 Posts: 5	Nanopore library on top of an Illumina library Hi all, we don't have the original aliquote of an old sample, but we have an Illumina Nextera Flex library in the freezer. I am aware that the ends are y-shaped, correct? Anyone of you have a fancy protocol to remodel/fill/modify those ends that I can do a Nanopore library on top? Thx!

11-30-2020, 09:45 AM	#2
luc Senior Member Location: US Join Date: Dec 2010 Posts: 452	No remodeling needed. All Nextera libraries are PCR amplified to be sequenceable. After the PCR amplification they are wonderfully double-stranded throughout and should be blunt-ended (if a high-fidelity polymerase was used). BTW, Truseq-style libraries have Y-adapters; Nextera/transposase libraries work differently.