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Bisulfite sequencing

Bisulfite sequencing is a collective name for a group of methods, aimed for DNA methylation pattern studies. Bisulfite treatment of DNA turns cytosine residues into uracil, without affecting 5-methylcytosine. After PCR amplification uracil is substituted by thymine, thus further analysis distinguishes single-nucleotide polymorphisms separating cytosines and thymines.

Comparison of the sequence obtained from the bisulfite-treated amplicon to the untreated one (published sequence) enables the identification of differential methylations.

Application examples:

Whole-genome methylation analysis.

Terms and prices:

Genome size Average coverage Run time Price per sample
3Gbx306-12 weeksRequest a quote
2Gbx306-12 weeksRequest a quote
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Purified DNA:

  • total amount — not less than 5 ug
  • concentration — not less than 100 ng/ul
  • OD260/280=1.8-2.0
  • no RNA contamination

Additional lab services:

  • Biomaterial-based DNA extraction
  • Repeated sample quality control
  • RNase treatment

Additional data analysis:

Basic bioinformatic services:
  • Sequencing quality and coverage uniformity tests
  • Data filtering (adapters and low-quality reads removal)
  • Reference reads mapping
Methylation analysis:
  • Methylation level detection
  • Promoters and CpG-islands coverage analysis
  • Identification of deferentially methylated regions (for 2 and more samples)