Library prep with sheared ssDNA

dtm2451 · 09-04-2013, 07:52 AM

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, 07:16 PM

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, 11:04 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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09-04-2013, 07:52 AM	#1
dtm2451 Junior Member Location: San Francisco, Ca Join Date: Mar 2013 Posts: 9	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.

09-04-2013, 07:16 PM	#2
Genohub Registered Vendor Location: genohub.com Join Date: Mar 2013 Posts: 210	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

02-19-2020, 11:04 AM	#3
BioDynami Registered Vendor Location: Alabama Join Date: Jul 2016 Posts: 23	There is a kit for ssDNA: NGS Single Stranded DNA Library Prep Kit from BioDynami: https://www.biodynami.com/ngs_single...brary_prep_kit