scAba-seq
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.
Advantages:
- Determines strand-specific 5hmC marks in individual cells
Disadvantages:
- Ambiguity can exist in 13% of the cleaved molecules (Serandour et al., 2016)
- Sequence bias in AbaSI activity (Sun et al., 2015)
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
None available yet
References:
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