SEQanswers

Go Back   SEQanswers > Applications Forums > Sample Prep / Library Generation



Similar Threads
Thread Thread Starter Forum Replies Last Post
Mixing cDNA samples from Ovation RNA‐Seq System V2 of NuGEN with Nextera Dual Indexes seq_uestion Sample Prep / Library Generation 1 11-19-2019 07:34 AM
Combining/multiplexing Nextera and XT indexes auzzie599 Illumina/Solexa 4 04-04-2019 06:22 AM
Using Nextera Indexes with Nextera XT Kit Angus's Microbiota Illumina/Solexa 7 11-11-2016 11:06 PM
Pooling and diluting Nextera libraries kmkocot General 0 09-17-2013 06:00 AM
Diluting Library for One Touch cwachtel Ion Torrent 1 04-23-2012 12:33 PM

Reply
 
Thread Tools
Old 10-15-2020, 08:41 AM   #1
Amygdala
Junior Member
 
Location: United Kingdom

Join Date: Sep 2019
Posts: 1
Default Diluting Nextera XT Indexes?

Hey all,

My samples originate from LCM, and I am currently following the SMART-seq2 workflow. Recently one group has combined both these approaches and written a nice protocol for it: https://doi.org/10.1007/978-1-4939-7213-5_6

Both Picelli's SMART-seq2 protocol (for single cells) and the above LCM-seq protocols mentions diluting the index adapters from the Nextera XT Index Kit, which appears to go against Illumina's protocol for adding 5ul of each adapter per sample without diluting. In the Picelli's original paper the authors do not dilute the index adapters, so I am a bit confused.

Can anyone clear this up for me? Or explain the benefit of diluting the adapters? I would have thought that since I am adding the recommended 1ng of input material for the Nextera XT library prep kit that I would follow Illumina's protocol of not diluting the adapters.

Many thanks!
Amygdala is offline   Reply With Quote
Reply

Thread Tools

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off




All times are GMT -8. The time now is 02:01 PM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2020, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO