SSB-Seq
SSB-Seq
SSB-Seq maps in vivo DNA single-strand breaks (SSBs) on a genome-wide scale. In this method, gDNA is isolated and subjected to nick-translation by DNA polymerase I in the presence of deoxynucleotide triphosphates (dNTPs), including digoxigenin-labeled dUTP, and a small amount of dideoxynucleotides. The DNA is sheared, and the labeled fragments are immunoprecipitated with antidigoxigenin antibodies and purified to prepare a sequencing library.
Pros:
- Simple protocol
Cons:
- The locations of the breaks cannot be determined accurately
- Not yet adopted widely by the scientific community