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. Author manuscript; available in PMC: 2014 Aug 20.
Published in final edited form as: Anal Chem. 2013 Aug 5;85(16):10.1021/ac400729q. doi: 10.1021/ac400729q

Figure 1.

Figure 1

Illustration of RT-LDR/spFRET assay for the expression analysis of mRNA transcripts. The assay utilizes no PCR and instead, employs a linear amplification scheme that can provide favorable limits-of-quantitation with high analytical sensitivity due to the digital readout format (i.e., single-molecule counting). Following reverse transcription, flanking primer pairs targeting unique sequences in specific mRNAs are subjected to ligation via the LDR. Successful ligation events generate MBs that undergo FRET. Single-molecule detection of MBs can give a direct readout of the mRNA copy number.