SF2-L1544 type
Library preparation is performed by using BGI Optimal DNA Library Prep Kit (BGI-Shenzhen China). A certain amount of genomic DNA is taken and subjected to fragmentation processing. Fragmented samples are size selected through magnetic beads. Then the size selected DNA fragments are converted to blunt-end DNA with end repair reaction. A single adenosine is added on the 3 end of DNA through the A tailing reaction. Later the library adaptors are connected to the two ends of DNA by adaptor ligation. Finally, the library products are amplified through PCR reaction and subjected to quality control process.Next, the final double strand library products are denatured to generate the single stranded library product. Then, the circularization reaction is set up to get single stranded circularized DNA products. Any single strand linear DNA will be digested to remove. The final single strand circularized library is amplified with phi29 and rolling circle amplification (RCA) to generate the DNA nano ball (DNB) which carries about 300 copies of the initial single stranded library molecule. The DNBs are loaded into the patterned nanoarray and sequencing reads of PE150 bases length are generated with DNBSEQ-G400 platform (BGI-Shenzhen, China)