First Strand cDNA Synthesis: Do not cool the mastermix containing E1 (step 3) and have the RNA / Custom Targeted Primer or RNA / FS1 / dT samples at 42 – 50 °C when adding the E1-containing mastermix (step 4) to avoid mishybridization. Mix properly by pipetting. Do not forget to shortly spin down the samples at room temperature before and after adding the E1-containing mastermix.
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