Detect 5hmC Marks in Single Cells With AbaSI Nuclease
scAba-seq is a single-cell, genome-wide, and strand-specific 5hmC sequencing technology (Mooijman et al., 2016). 5hmC marks in DNA from individual cells are glucosylated with T4 phage _-glucosyltransferase (T4-_GT_, and the DNA is digested with the restriction endonuclease AbaSI. The digested DNA is ligated to an adapter containing a cell-specific barcode, an Illumina 5_ adapter, and a T7 promoter. The ligated DNA from different cells is pooled and amplified using in vitro transcription mediated by T7 RNA polymerase.
- Determines strand-specific 5hmC marks in individual cells
- Ambiguity can exist in 13% of the cleaved molecules (Serandour et al., 2016)
- Sequence bias in AbaSI activity (Sun et al., 2015)
Illumina Library prep and Array Kit Selector
None available yet
Mooijman D., Dey S. S., Boisset J. C., Crosetto N. and van Oudenaarden A. Single-cell 5hmC sequencing reveals chromosome-wide cell-to-cell variability and enables lineage reconstruction. Nat Biotechnol. 2016;34:852-856