Hairpin Priming Is Better Suited than In Vitro Polyadenylation to Generate cDNA for Plant miRNA qPCR  Sajag Adhikari, Marie Turner, Senthil Subramanian  Molecular Plant  Volume 6, Issue 1, Pages 229-231 (January 2013) DOI: 10.1093/mp/sss106 Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 1 Comparison of Hairpin and polyA cDNA qPCR to Assay Synthetic RNA Molecules With and Without 2’-O Methylation, Selected miRNAs from Methylation-Deficient Arabidopsis hen1 Mutants and miR164 along a Time Course of Rhizobium Inoculation in Soybean Indicates that Hairpin cDNA qPCR Is Better Suited to Assay Plant miRNAs.(A–D) Amplification plots from hairpin (A, C) and polyA (B, D) cDNA qPCR assays to detect different amounts of 2’-OH and 2’-OMe RNA molecules (1 fmole, blue circles; 0.1 fmole, red squares; 0.01 fmole, green triangles; and 0.001 fmole, gray diamonds).(E) Expression of miR160, 164, 166, and 393 in Arabidopsis wild-type and hen1 seedlings assayed by hairpin cDNA qPCR (shaded bars) and polyA cDNA qPCR (black bars). Data shown are the ratio of normalized miRNA abundance in hen1 over wild-type and representative of independent experiments. In all cases, expression in hen1 was significantly different from that in WT (P < 0.01; Student’s t-test).(F, G) Relative abundance of miR164 (versus 0 h time point) assayed by Northern hybridization (blue diamonds), hairpin cDNA qPCR (red squares), and polyA cDNA qPCR (green triangles) along a time course of 14 d in (F) mock- and (G) B. japonicum-inoculated soybean roots. Detailed methods in supplementary information. Molecular Plant 2013 6, 229-231DOI: (10.1093/mp/sss106) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions