Table 1.
On-target efficiency for libraries prepared from untreated RainDance captured DNA.
Figure 1.
Mung bean nuclease treatment abolished the high molecular DNA smear in RainDance captured DNA after end repair.
A. An overview of the workflow. Aliquots of 100ng of DNA enriched through microdroplet-PCR were either treated with mung-bean nuclease or untreated as a control. The differentially treated aliquots were end-repaired, concatenated, processed into TruSeq libraries and sequenced on MiSeq in parallel to rule out any batch effect. B. Electrophoresis analysis of DNA samples on a high-sensitivity DNA chip using the Agilent 2100 Bioanalyzer. DNA samples in all 4 lanes were derived from 200pg of the same batch of captured DNA. From left to right, the lanes are, “L” lane-the DNA size ladder, lane 1-untreated DNA enriched by RainDance microdroplet-PCR prior to end repair, lane 2- untreated DNA enriched by RainDance microdroplet-PCR post end repair, lane 3- mung bean nuclease treated DNA enriched by RainDance microdroplet-PCR prior to end repair, lane 4- mung bean nuclease treated DNA enriched by RainDance microdroplet-PCR post end repair.
Figure 2.
Mung bean nuclease treatment significantly increases on-target efficiency for DNA enriched through RainDance microdroplet-PCR.
Aliquots of RainDance enriched DNA for the same sample were either treated or not treated with mung bean nuclease, processed into TruSeq libraries, and sequenced on MiSeq as illustrated in Figure 1A. Plotted is the mean value of on-target efficiency of 3 samples that went through parallel treatments (also see Table 2). Nuclease treatment leads to significantly higher on-target efficiency (*p = 0.018, one-tail paired t test; error bar, SEM).
Table 2.
On-target efficiency for libraries prepared from samples either treated or untreated with the mung bean nuclease.
Table 3.
Comparison of variants detected for the same sample either treated or not treated with mung bean nuclease.