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Fig. 4 | BMC Genomics

Fig. 4

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

Fig. 4

Comparison of PCR approaches in terms of proportion of chimeric products generated per BC from MPRA libraries. BC–ROI regions of library-71 and library-83 were amplified by emulsion and conventional two-round PCR using the same settings (10 pg of template DNA, 15 amplification cycles and elongation time of 30 s in round #1 PCR, 1/100th of the purified products of round #1 PCR as template, 18 amplification cycles and elongation time of 30 s in round #2 PCR) and subsequently subjected to NGS. The experiments were done in duplicates and averaged values of proportion of chimeric products per BC are plotted as histograms separately for library-71 (a) and library-83 (b). Pronounced peaks at middle values of proportion of chimeric products per BC are mainly a result of low NGS coverage of some BCs (i.e., the peaks primarily represent cases with 1 chimeric BC–ROI combination per several genuine ones)

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