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Table 1 Occurrence of chimeric BC–ROI combinations under different ePCR conditions for the two-plasmid template system

From: Optimized PCR conditions minimizing the formation of chimeric DNA molecules from MPRA plasmid libraries

Round #2 ePCR conditions Total read count Reads with chimeric BC–ROI combinations, %
R#1 R#2 BC1–ROI2 BC2–ROI1 Average
BC1 BC2 BC1 BC2 R#1 R#2 R#1 R#2 R#1 R#2
0.5 μl of round #1 ePCR products, 20 cycles 134,011 134,779 47,072 46,333 1.579 1.651 1.465 1.347 1.522 1.499
0.3 μl of round #1 ePCR products, 20 cycles 43,782 44,788 10,327 10,452 0.831 0.629 0.737 0.440 0.784 0.535
0.5 μl of round #1 ePCR products, 18 cycles 21,042 21,733 166,862 170,934 0.200 0.225 0.230 0.211 0.215 0.218
  1. The settings of the round #1 ePCR were the same for all samples: 15 amplification cycles starting with ~ 2 × 106 template DNA molecules
  2. R#1 and R#2 stand for replicate 1 and replicate 2, respectively