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-   -   Library prep with sheared ssDNA (http://seqanswers.com/forums/showthread.php?t=33413)

dtm2451 09-04-2013 06:52 AM

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.

Genohub 09-04-2013 06:16 PM

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

BioDynami 02-19-2020 10:04 AM

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


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