Neurospora crassa
Qualified qualified DNA sample(s) was constructed BS library. Purified DNA sample, such as genomic DNA is sheared into smaller fragments with a desired size by Covaris S/E210 or Bioruptor firstly. Then the overhangs resulting from fragmentation were converted into blunt ends by using T4 DNA polymerase, Klenow Fragment and T4 Polynucleotide Kinase. After adding an A base to the 3' end of the blunt phosphorylated DNA fragments, adapters are ligated to the ends of the DNA fragments. The desired fragments were purified though gel-electrophoresis, then selectively enriched and amplified by PCR. The index tag was introduced into the adapter at the PCR stage as appropriate and we did a library quality test. At last, the qualified BS library was used for sequencing.