SSB-Seq
Map DNA Single-Strand Breaks
SSB-Seq maps in vivo DNA single-strand breaks (SSBs) on a genome-wide scale (Baranello et al., 2014).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.
Advantages:
- Simple protocol
Disadvantages:
- Locations of breaks cannot be determined accurately
- Not yet adopted widely by the scientific community
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
None available yet
References:
Baranello L., Kouzine F., Wojtowicz D., et al. DNA break mapping reveals topoisomerase II activity genome-wide. Int J Mol Sci. 2014;15:13111-13122