I am setting up high-throughput miRNA profiling experiments. What is the typical workflow for these experiments?
A typical workflow is shown in the figure “miRNA profiling workflow”. Steps in a typical workflow are as follows. Purify total RNA from the samples of interest. RNA preparations must contain small RNAs (e.g., RNA prepared using miRNeasy Kits includes RNA from approximately 18 nucleotides upwards). A typical experiment uses RNA from 2 samples: a control sample (e.g., untreated, normal) and a test sample (e.g., treated, disease). A positive control, such as a synthetic, single-stranded miRNA, may also be included. Dispense miScript Primer Assays from the source plate into a 384-well PCR plate. In a typical experiment, each PCR will be performed in triplicate. This means that each miScript Primer Assay will be dispensed into 9 wells (3 samples [control, test, synthetic miRNA] x 3 replicates). Perform a reverse-transcription reaction using the miScript Reverse Transcription Kit with the purified total RNA as template. Dilute and divide the reaction into aliquots, depending on how many miRN
