Supplementary Figure 1: A new qRT-PCR approach suitable for quantifying any small RNAs.

(a) Overview of the approach. The sRNA sample is ligated to a 3’ DNA linker, which then acts as a priming site for cDNA synthesis primer. An additional 5’ RNA adapter is ligated to the 3’ linker ligated RNA sample, prior to cDNA synthesis. After cDNA synthesis, two options are available for specific qPCR of the target sRNA. Each option uses a primer specific to the sRNA sequence and another primer designed to bind to either the 3’ adapter or the 5’ adapter, allowing flexibility in primer design. (b) Example of the application of this qPCR method for the miR29b1 as a part of quality checking during the TEsR procedure. A universal reverse primer and a forward primer specific to miR29b1 were used. Presence of qRT-PCR products was confirmed by agarose gel, and quantitative enrichment of the library was confirmed by qPCR of mir29b1 in enriched and unenriched libraries.