Skip to main content
. 2020 Jun 16;48(14):e84. doi: 10.1093/nar/gkaa500

Figure 1.

Figure 1.

Preparing and evaluating synthetic RNA–DNA hybrids as spike-ins for DRIP. (A) Experimental scheme showing how hybrids were synthesized. Briefly, target regions were amplified from E. coli genomic DNA with a flanking T7 promoter. RNA was prepared from these templates by in vitro transcription, then hybridized to a synthetic ssDNA oligo. Hybrids were purified by agarose gel electrophoresis. (B) Gel image showing RNase H reversible size-shifts after hybridization of RNA and DNA. Unlabeled samples were separated on a 2.5% agarose gel which was then stained with RedSafe nucleic acid staining solution. (C) qPCR of genomic (left) and spike-in (right) hybrids following transcription inhibition with DRB. RNase H (RH) treatment demonstrates antibody specificity. Error bars represent 95% confidence interval (CI) of the mean. Results are significantly different as determined by non-overlapping 95% CIs. In primer name, GB indicates gene body.