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Figure 3 | Molecular Cancer

Figure 3

From: Optimized high-throughput microRNA expression profiling provides novel biomarker assessment of clinical prostate and breast cancer biopsies

Figure 3

Schematic of microRNA amplification procedure. A poly A tail is first added at the 3' end of microRNAs. Reverse transcription performed using a primer consisting of an oligo dT and a capture sequence. Next, the first strand cDNA is tailed with dTTP using terminal deoxynucleotidyl transferase on the 5' end, followed by annealing of a T7 template oligo to the 3' tail of the cDNA. Klenow enzyme fills in the 3' end of the first strand cDNA to produce a double-stranded T7 promoter. The T7 template contains a blocker to prevent second strand synthesis. An in vitro transcription reaction with T7 RNA polymerase is then performed. Amplified products are then labeled using the method previously mentioned starting at the reverse transcription step.

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Molecular Cancer

ISSN: 1476-4598