SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
Making library from edge of sheared DNA smear - possible biases? Noa Illumina/Solexa 1 08-21-2013 07:49 AM
Possible bias from making library from edge of Covaris-sheared DNA? Noa Bioinformatics 0 08-18-2013 10:42 AM
Illumina library construction from ssDNA (MeDIP)?? lihzhou Sample Prep / Library Generation 2 07-27-2013 04:42 AM
custom ssDNA library with strange size distribution of qPCR products pyridine Sample Prep / Library Generation 4 07-17-2013 03:18 AM
Library prep woes unusual bioanalyzer profile of Illumina RNA-seq library petey Sample Prep / Library Generation 1 11-19-2012 09:00 AM

Reply
 
Thread Tools
Old 09-04-2013, 06:52 AM   #1
dtm2451
Junior Member
 
Location: San Francisco, Ca

Join Date: Mar 2013
Posts: 9
Question Library prep with sheared ssDNA

Does anyone know the integrity/makeup of the 5' and 3' ends of Covaris-sheared ssDNA? I am working on a library prep method for single stranded cDNA and am hoping to ligate adapters post shearing.

My assumption is that the cleavage could not leave intact 5' phosphate and 3' OH. Therefore, I am also wondering about methods to repair these ends that would not require generation of a second strand.
dtm2451 is offline   Reply With Quote
Old 09-04-2013, 06:16 PM   #2
Genohub
Registered Vendor
 
Location: genohub.com

Join Date: Mar 2013
Posts: 210
Default ssDNA ligation

Check out this article, it should help: Single-stranded DNA library preparation for the sequencing of ancient or damaged DNA.

If you don't follow the approach in the paper, you can use T4 Polynucleotide Kinase to add a 5'- Phosphate and remove 3-phosphoryl groups.


- Genohub
Genohub is offline   Reply With Quote
Old 02-19-2020, 10:04 AM   #3
BioDynami
Registered Vendor
 
Location: Alabama

Join Date: Jul 2016
Posts: 22
Default

There is a kit for ssDNA: NGS Single Stranded DNA Library Prep Kit from BioDynami:
https://www.biodynami.com/ngs_single...brary_prep_kit
BioDynami 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 12:19 AM.


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