Fig. 2
From: DeLTa-Seq: direct-lysate targeted RNA-Seq from crude tissue lysate
Quantification of cDNA synthesized by direct-lysate reverse transcription. a qPCR results for the Os03g0836000 transcript in the cDNA synthesized from mixtures of purified O. sativa total RNA and the reducing reagents dithiothreitol (DTT) and 2-mercaptoethanol (2ME). The horizontal axis indicates the concentrations of the reducing regents in the RT reaction mixes. The vertical axis indicates the differences in Ct value compared to the RT(−) negative control. Each point indicates a value of each replicate. Each horizontal line indicates the average of each condition. b–e qPCR results for AT3G18780 transcript (b), Os03g0836000 transcript (c, d), and HP620998.1 transcript (e) in cDNA synthesized from lysate of A. thaliana seedlings (b), the youngest fully-expanded leaves of O. sativa (c), O. sativa roots (d), and T. aestivum coleoptile (e). The horizontal axis indicates the concentrations of the reducing reagents in the homogenization buffers. The vertical axis indicates the differences in Ct value compared to the RT(−) negative control. Each point indicates a value of each replicate. Each horizontal line indicates the average of each condition. Same amount of total RNA was used as cDNA template according to RNA concentrations in lysates (250 ng of A. thaliana and O. sativa or 140 ng of T. aestivum). f A schematic diagram of the primer sets for cDNA detection. g qPCR results for Os03g0836000 transcript in cDNA synthesized from lysate of O. sativa leaves. In the direct-lysate reverse transcription, 0.1, 0.5, 1, 2.5, 5, 10 and 12.5 µL of the lysate were used. Three technical replicates were prepared. The red line is a linear regression line