Ultrafast bisulfite sequencing detection of 5-methylcytosine in DNA and RNA

Abstract Bisulfite sequencing (BS-seq) to detect 5-methylcytosine (5mC) is limited by lengthy reaction times, severe DNA damage, overestimation of 5mC level and incomplete C-to-U conversion of certain DNA sequences. We present ultrafast BS-seq (UBS-seq), which uses highly concentrated bisulfite reagents and high reaction temperatures to accelerate the bisulfite reaction by ~13-fold, resulting in reduced DNA damage and lower background noise. UBS-seq allows library construction from small amounts of purified genomic DNA, such as from cell-free DNA or directly from 1 to 100 mouse embryonic stem cells, with less overestimation of 5mC level and higher genome coverage than conventional BS-seq. Additionally, UBS-seq…

• NS
  National Science Foundation

  Awards: 1048528, CHE-1048528
• UO
  University of Chicago
• NI
  National Institutes of Health

  Awards: HG008935, R01 HG006827, RM1 HG008935, HG006827
• DO
  Division of Chemistry

  Award: CHE-1048528