cool to either 58 °C. In all, 14 μl of extension mix (0.12 μl of avian myeloblastosis virus reverse transcriptase (Promega), 2.8 μl 5X RT buffer, 1.12 μl 1 mM deoxynucleotidetriphosphates, RNase-free water to 14 μl) was added to each reaction and incubated at 58 °C for 1 h. Samples were then mixed with 2X formamide denaturing dye, heated to 95 °C for 3 min and resolved on an 18% polyacrylamide, 7 M urea denaturing gel. Gels were exposed by Phosphor-Imager analysis. Positive controls underwent same treatment but started with 3.2 μg of RNA.
