Library prep with sheared ssDNA

[dtm2451]

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]

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]

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