How to construct a sgRNA library with multiple samples?
0
1
Entering edit mode
2.2 years ago
frymor2 ▴ 10

I was wondering if anyone has already experience with constructing sgRNA libraries. We are interested in doing a knock-out experiment (loss-of-function) with 5 different conditions. I have found this protocol from Joung et al.

In the protocol they have 10 different Fwd primer with no unique barcodes, only the eight Rev primers contains unique barcodes for different samples in KO.

Do I understand it correctly, that i can pool together only eight samples in one sequencing run with this protocol?

Are there more complex library preparation kits out there which can deal with more samples?

thanks

sgRNA library crispr-cas9 NGS • 492 views
ADD COMMENT
1
Entering edit mode

This questions is best asked on SeqAnswers.com or biology stackexchange. One part of your question can be answered here:

Are there more complex library preparation kits out there which can deal with more samples?

There are kits available with unique single and dual indexes that allow hundreds of sample libraries to be pooled together and run in one lane for Illumina. They are then demultiplexed to get individual sample files.

ADD REPLY

Login before adding your answer.

Traffic: 1490 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6