Journal: bioRxiv
Article Title: Development of Coupling Controlled Polymerizations by Adapter-ligation in Mate-pair Sequencing for Detection of Various Genomic Variants in One Single Assay
doi: 10.1101/400689
Figure Lengend Snippet: The first adapter (Ad1, blue) with/without a barcode sequence (indicated by grey bar) is ligated to genomic DNA fragments (black lines). Various adapter designs may be used as the first adapter [e.g., blunt-end adapters, Y-shaped Illumina adapters, bubble adapters ( Supplementary Fig. S2 ), etc.]. After Ad1 ligation and PCR, DNA ends are ligated together to form dsCirs containing a gap. naCNT is performed and results in DNA polymerization (dashed lines) and the movement of the gap into a selected length of the genomic DNA. 3’-branch ligation is used to ligate a 3’-end of the second adapter (Ad2_3’, yellow). The two strands of the dsCir are separated, and the single-stranded DNA (ssDNA) with Ad2_3’ at the 3’-end is used as a template for CPE (ntCPE or ttCPE). The Ad2_5’ sequence (green) is added to the 3’-end of the CPE product through 3’-branch ligation. After PCR, this results in genomic DNA with half of Ad2 at each end separated by Ad1.
Article Snippet: For all four libraries, single-stranded circles (ssCirs) and DNA nanoballs (DNBs) were prepared and subjected to improved cPAL (Combinatorial Probe-Anchor Ligation) sequencing on a DNA nanoball (DNB)-based sequencing platform [Complete Genomics, Inc., (CGI), Mountain View, CA, USA].
Techniques: Sequencing, Ligation