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Prepare siRNA and miRNA Probes in Just 1 Hour

eed to supply is a short, inexpensive DNA oligonucleotide specific for the target of interest that also includes an 8 base sequence complementary to the 3' end of the T7 promoter primer included in the kit. This oligonucleotide is annealed to the provided T7 promoter primer, and a 30 minute fill-in reaction with Klenow fragment generates the double-stranded transcription template. The resulting template is simply mixed with the provided T7 RNA Polymerase, rNTPs, and Transcription Buffer along with your choice of labeled rNTP. The reaction is then incubated for as little as 10 minutes at 37C to generate labeled probe .


Complete Kit for Probe Preparation
The kit includes enough reagents to prepare 30 dsDNA templates and to perform 30 in vitro transcription reactions. Each template construction reaction supplies sufficient template for up to 20 transcription reactions. The kit also includes a functional positive control, DNase I to remove residual DNA template, and Probe Elution Buffer to gel purify RNA transcripts.

Figure 1. miR-16 and miR-22 Expression in Mouse Brain. In situ hybridization was performed on coronal mouse forebrain sections. In situ hybridized cells in the mouse brain cortex (A,B,D,E; 20X magnification) and the head of the caudate nucleus (C,F; 40X magnification) are indicated by arrows. Nonisotopically labeled probes were prepared with Ambion's MAXIscript Kit (1.5 kb probe) or with the mirVana miRN
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