Article Title: COBRA-Seq: Sensitive and Quantitative Methylome Profiling
Figure Lengend Snippet: GW-COBRA and LA-COBRA library construction results. ( A ) Sonicated genomic DNA isolated from HCT116 cell line (lane 1), 50 ng of Adapter-2 ligated library material (lane 2). Column purified Adapter-2 ligated library material (lane 3); ( B ) PCR amplification test of bisulfite treated library materials. Lane 1, 2, 3 and 4 are produced by 6, 7, 8 and 9 cycles of PCR using GW-A2-FwdP and COBRA-A2-RevP primers from Table 1 with GW-COBRA library material as the template respectively. Lane 5, 6, 7 and 8 are produced by 6, 7, 8 and 9 cycles of PCR using LA-A2+T7-FwdP and COBRA-A2-RevP primers from Table 1 with LA-COBRA library material as the template, respectively; ( C ) Final library products: GW-COBRA and LA-COBRA methylome libraries of HCT116 DNA amplified using flowcell or LADS primer pairs respectively (Lane 2 and 4). Lane 1 and 3 are negative controls for GW-COBRA and LA-COBRA libraries, respectively; ( D ) Bioanalyzer results of GW-COBRA and LA-COBRA methylome sequencing libraries, respectively, prepared from HCT116 cell line DNA. The final library fragments ranged between 150–500 bp with an average size of 257 and 360 bp for GW-COBRA and LA-COBRA, respectively.
Article Snippet: Finally, the size distribution was visualized using Agilent DNA 1000 Assay in 2100 Bioanalyzer (Agilent Technologies, Los Angeles, CA, USA) using the manufacturer’s protocol.
Techniques: Combined Bisulfite Restriction Analysis Assay, Sonication, Isolation, Purification, Polymerase Chain Reaction, Amplification, Produced, Sequencing