genome sequencing of Bacillus coagulans
Genomic DNA of B. coagulans Na-2 was extracted and sheared into smaller fragments with a desired size by Covaris S/E210 or Bioruptor firstly. Then the overhangs resulting from fragmentation are 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 the quality of the library was tested. At last, the qualified BS library was used for sequencing. Sequencing was carried out on the Illumina HiSeq 2000 platform (Beijing Genomics Institute, Beijing, China).