Technical Considerations for Reduced Representation Bisulfite Sequencing with Multiplexed Libraries

Figure 1

Representative analytical PCR of size-selected RRBS libraries.The 160–340 bp size-selected RRBS libraries (represented by a, b, and c) were amplified with either 15 or 20 cycles of PCR to determine the optimal number of cycles for large-scale amplification. PCR products were visualized on a 4–20% Criterion gradient polyacrylamide TBE gel stained with SYBR green nucleic acid gel stain alongside a 25 bp DNA ladder. For these libraries, 13 (a) and 14 (b and c) cycles were chosen for large-scale amplification. The distinct band at 125 bp in all libraries was possibly due to adaptor-adaptor dimerization.