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Detecting Attomole Amounts of Small RNA

A probes and the included Hybridization Buffer. After heat denaturation, each mixture is incubated at 42C to hybridize the probe to its complementary RNA molecules. Unhybridized RNA species and excess RNA probe is then removed by a rapid ribonuclease digestion step. The hybridized, protected RNA fragments are recovered in the same tube using Ambion's patented single step technology for simultaneous ribonuclease inactivation and nucleic acid precipitation. RNA samples are then resuspended with the provided Gel Loading Buffer and analyzed on a denaturing polyacrylamide gel. Because the hybridization is performed in solution, the procedure ensures a sensitive and linear detection signal after autoradiography. Furthermore, the included Hybridization Buffer has been specifically developed to provide the optimal sensitivity and specificity of detection with short antisense probes. Thus the procedure is ideal for detecting small RNA molecules such as siRNA or miRNA.


Complete Kit with Included Controls
Each mirVana miRNA Detection Kit provides enough reagents for 100 reactions. The kit also contains a control transcription template and Probe Elution Buffer to prepare and gel purify a 32 nt RNA probe specific for miR-16 miRNA. When used with the provided Mouse Kidney Total RNA, this probe generates a 22 nt protected fragment. The kit also comes with a detailed Instruction Manual and comprehensive information about experimental setup and probe design/preparation.

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