Acoustic microstreaming increases the efficiency of reverse transcription reactions comprising single-cell quantities of RNA

Abstract

Correlating gene expression with behavior at the single-cell level is difficult, largely because the small amount of available mRNA (<1 pg) degrades before it can be reverse transcribed into a more stable cDNA copy. This study tested the capacity for a novel acoustic microstreaming method ("micromixing"), which stirs fuid at microliter scales, to improve cDNA yields from reverse transcription (RT) reactions comprising single-cell quantities of RNA. Micromixing significantly decreased the number of qPCR cycles to detect cDNA representing mRNA for hypoxan-thine phosphoribosyl-transferase (HPRT) and nuclear receptor-related 1 (Nurr1) by ~9 and ~15 cycles, respectively. Te improvement was equiva..