Illumina_DNASeq_DZ2_Genome
Sample testing included concentration, sample integrity and purity. Concentration was detected by fluorometer or Microplate Reader (e.g. Qubit Fluorometer, Invitrogen). Sample integrity and purity were detected by Agarose Gel Electrophoresis (Concentration of Agarose Gel:1% Voltage:150 V, Electrophoresis Time:40 min). 1g genomic DNA was randomly fragmented by Covaris. The fragmented genomic DNA were selected by Magnetic beads to an average size of 200-400bp. Fragments were end repaired and then 3 adenylated. Adaptors were ligated to the ends of these 3 adenylated fragments. This process was to amplify fragments with adaptors from previous step. PCR products were purified by the Magnetic beads . The double stranded PCR products were heat denatured and circularized by the splint oligo sequence. The single strand circle DNA (ssCir DNA) were formatted as the final library. Library was qualified by QC. The library was amplified with phi29 to make DNA nanoball (DNB) which have more than 300 copies of one molecular. The DNBs were load into the patterned nanoarray and pair end 100/150 bases reads were generated in the way of combinatorial Probe-Anchor Synthesis (cPAS).