Available Technologies

Background

Methylation of DNA is one of epigenetic mechanisms and is deeply associated with embryo generation, cell differentiation, phenotype differences, various diseases, and the like. Further, methylation analysis of DNA is also used for sorting iPSC. In recent years, attempts are made to perform methylation analysis of single cell DNA using RRBS (Reduced Representation Bisulfite Sequencing) method capable of examining the methylation of DNA at a single base level. However, the DNA sample is fragmented because the bisulfite treatment is performed under a severe condition, and sufficient amount of DNA for PCR amplification is not supplied for further amplification for the next step.

Description and Advantages

Performing rescue including a step of treating a DNA sample after bisulfite treatment with single-stranded DNA ligase solves such problems, and can perform PCR amplification after bisulfite.
Rescue reaction The rescue reaction included following four steps:
1) Immobilization on beads
2) Phosphatase reaction
3) Kinase reaction
4) Single-stranded DNA ligase reaction