A universal TaqMan-based RT-PCR protocol for cost-efficient detection of small noncoding RNA

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FIGURE 5.
FIGURE 5.

Detection of artificial and endogenous small noncoding RNAs in prokaryotes and eukaryotes with the universal probe. (A) Confirmation of delivery of siRNA after electroporation of Listeria monocytogenes compared with buffer and control siRNA. Relative values normalized to the Listeria 6PBG housekeeping gene and compared with unpulsed samples (2−(ΔΔCT) method). Errors represent standard deviation from ΔΔCT, one experiment in duplicate. (B) Detection of two small noncoding RNAs CsrB and Lst-R isolated from Escherichia coli grown in log phase in rich media. (NTC) No template control. (C,D) Detection of endogenous miR-133b and miR-146a using the universal design (Com) and universal modified (Mod) protocol in total RNA from human cell lines HEK293T and Jurkat. Error bars represent value range from mean of one experiment performed in duplicate.

This Article

  1. Published in Advance October 22, 2013, doi: 10.1261/rna.040501.113 RNA 19: 1864-1873

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