Remarkable stability of mRNA and miRNA at repeated freeze-thaw cycles in the <i>in-house</i> lysates.

<p>M14 cultured in 96-wells at 10⁴ cells per well (biological triplicates) were directly lysed by (A) in-house or (B) Bio-Rad lysis buffers. Cell lysates were collected and aliquoted. The lysates were then frozen at −70°C, thawed in a temperature-controlled water bath at 25°C, and the freeze-thaw cycles repeated. The miRNA and mRNA expressions were quantified following 1, 3, 5, 10 freeze-thaw cycles. The qPCR assays from set 2 were used for quantifications. Ct was plotted against freeze-thaw cycles to examine the stability of RNA. The experiments were conducted in biological duplicates.