RT² miRNA First Strand Kit

The RT² miRNA First Strand Kit contains all reagents needed for converting miRNA into first strand cDNA for real-time PCR. The simple one-step reaction not only performs both poly-(A) tailing and reverse transcription at the same time. It also incorporates a proprietary and unique universal PCR priming sequence providing a scalable analysis solution for multiple miRNA sequences in the same cDNA preparation. The patent-pending miRNA First Strand formulation improves the specificity of reverse transcription for mature miRNA sequences over their precursors, primary transcripts, and mRNA targets. The kit also provides greater control over RNA quality with its built-in External RNA Control as well as enhanced sensitivity over other suppliers' miRNA first strand cDNA synthesis kits. Due to the unique nature of the universal PCR priming sequence, the RT² miRNA First Strand Kit is specifically designed for maximizing the performance of the RT² miRNA PCR Array and Assay Systems. Please click here for pricing & ordering information.

Benefits of the RT² miRNA First Strand Kit

• Improved Specificity & Sensitivity: Proprietary reaction conditions do not reverse transcribe miRNA precursors, primary transcripts, or mRNA - only mature miRNA is detected
• Insure RT Efficiency & Consistency: Built-in External RNA Control verifies lack of enzyme inhibitors and insures efficient reverse transcription
• Easy and Fast: Everything included in one kit to complete first strand cDNA synthesis in a one-step reaction

HOW IT WORKS:

1. A Poly-A Polymerase adds a poly-A tail to the miRNA sequence.
2. A primer, containing oligo(dT) and a universal PCR primer sequence, anneals to the poly-adenylated RNA.
3. A Reverse Transcriptase synthesizes and appends to the primer DNA sequence complementary to the miRNA.
4. The final first strand cDNA product contains the miRNA complement sequence, a poly(dT) tract, and the universal PCR primer sequence. It is immediately ready for a real-time PCR assay containing a miRNA-specific and the universal PCR primer.

Control:
A built-in miRNA External RNA Control (ERC) helps test for inhibitors of reverse transcription. Control elements on the RT² miRNA PCR Arrays detect template synthesized from the ERC. A reproducible threshold cycle value from this control across your samples indicates a consistently high level of RNA quality and transcription efficiency giving you a greater degree of confidence in your results.

Easy and Fast Sensitivity:
Combining poly(A) tailing and reverse transcription in one reaction and one tube saves time and sample while reducing variability. The optimized reaction conditions and buffer components maximize cDNA product yield using anywhere from 50 to 400 ng of input small RNA, and are most compatible with our RT² SYBR Green qPCR Master Mixes. The universal PCR priming sequence incorporated during reverse transcription also provides a scalable solution for analyzing the expression of multiple miRNA sequences in the same cDNA preparation under standardized conditions.

Complete RT² miRNA PCR System
Other sources of miRNA reverse transcription assay will not work with the RT² miRNA PCR Arrays and Assays, and cDNA from the RT² miRNA First Strand Kit cannot be used with other source of miRNA qPCR assays. Only the complete RT² miRNA PCR System yields detectable real-time PCR results.

For more information, see our RT² miRNA PCR Array System User Manual [here], or see the RT² miRNA First Strand Kit Specification Sheet [here].

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