SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
TruSeq PCR Free LT Kit Issues addyblanch Illumina/Solexa 4 04-26-2018 09:22 AM
TruSeq DNA PCR-Free Sample Preparation Kit (Illumina) expired 3 months - can I use? snarapor Sample Prep / Library Generation 3 08-22-2015 06:59 AM
Truseq DNA PCR-free kit Laruo Sample Prep / Library Generation 1 03-05-2014 03:42 AM
NEBNext ChIP-Seq library prep kit vs Illumina TruSeq kit? ASL Sample Prep / Library Generation 1 04-10-2013 08:35 PM
The NEXTflex™ PCR-Free DNA Sequencing Kit Reduces Bias in NGS Library Prep Bioo Scientific Vendor Forum 10 12-20-2012 08:10 AM

Reply
 
Thread Tools
Old 06-08-2016, 05:28 AM   #1
Lovro
Member
 
Location: Slovenia

Join Date: Mar 2015
Posts: 19
Default which library kit; are adapters on Illumina PCR free LT kit biotinylated ?

Hi,
I would like to prepare an enriched (custom IDT lockdown probes) NGS (Illumina platform) library that has a PCR step as late as possible in the protocol - I want to bisulfite treat the already adapter tagged and enriched library or part of it.
Ideally, I would be able to add adapters, use IDT probes, bisulfite treat 2/3 of the library, amplify with PCR, and then in 24 multiplex run the library on Illumina platform. I want to do this separately for bisulfite treated and non-treated parts of the library.
Input to protocol would be a Covaris sonicated human genome DNA, cut to 800 - 1000bp

For that, I guess, I would need a kit that has the following characteristics (please comment if you think they are unnecessary or perhaps unrealistic):

1. High DNA input (1-2 micro gram of starting DNA or more), since the enriched target is relatively small.
2. Adapters which have the cytosins methylated (to resist bisulfite), but are not biotinylated (to not be pulled by the enrichment streptavidin beads).
3. No PCR before I can bisulfite treat (right after enrichment step)

I see that Illumina TruSeq DNA PCR-Free LT
(http://support.illumina.com/sequenci...ee-lt-kit.html)
satisfies my needs, except I don't know if adapters are biotinylated.
It bothers me that in their webinar of kit comparison
(http://support.illumina.com/content/...ry.html?iframe )
the TruSeq DNA PCR-Free LT is marked as non enrichment-compatible.

QUESTION 1:
Does anybody know, if the adapters in TruSeq DNA PCR-Free LT are biotinylated?
QUESTION 2:
Does anybody know of any other NGS library prep kits that would satisfy my needs even better or for a lesser price?


Enrichment has to be before bisulfite treatment and as far as I know, IDT probes require the presence of adapters, so please don't suggest bisulfite treating before library preparation.

Thanks a million,
Lovro
Lovro is offline   Reply With Quote
Reply

Tags
adapter, bisulfite conversion, enrichment, ngs library prep, truseq dna pcr-free

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 04:31 PM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2018, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO