The microScript microRNA cDNA Synthesis Kit enables reverse transcription of microRNA from either total RNA or enriched microRNA preparations.
Size
- 50 reactions
- 12 reactions
Norgen’s microScript microRNA cDNA Synthesis Kit is a complete, ready-to-use kit for reverse transcription of microRNA from either whole RNA or enriched microRNA preparations.
The kit includes the 2× Reaction Mix and the microScript microRNA Enzyme Mix. The kit employs Norgen's microScript Reverse Transcriptase, a mutant form of Moloney Murine Leukemia Virus (M-MuLV) Reverse Transcriptase. It has lower RNase H activity and higher heat stability.
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Features and benefits
- Ease of use
- One cDNA Synthesis, Multiple microRNAs, and microRNA-targets analyzed
- Strong Enzyme
- Available in 12 or 50 reaction size
- Time Savings
- Cost Efficient
- High Sensitivity and Yield
Norgen’s microScript microRNA cDNA Synthesis Kit has a simple, single-tube setup that includes mixing 2× Reaction Mix, Enzyme Mix, and RNA template. The reaction can then be run in a thermocycler. A poly (A) tail is first added to the RNA template, and then cDNA is synthesized using an adaptor primer. In addition to its simplicity, the single-tube setup offers exceptional consistency and sensitivity.
A Universal PCR Reverse Primer and a forward primer containing the sequence of the target microRNA could be used to amplify the cDNA in a PCR or qPCR. Several distinct microRNAs could be amplified by PCR using a single cDNA preparation.
If total RNA preparation was the starting template, the cDNA preparation might be utilized for PCR or qPCR detection of mRNA or big RNA (using gene-specific forward and reverse primers). This would make it possible to assess the expression levels of microRNAs and microRNA-targets simultaneously.
Details
Supporting data
Figure 1. High Sensitivity with Robust Amplification of microRNAs from Different Amounts of RNA Input using Norgen’s microScript microRNA cDNA Synthesis Kit. Norgen’s microScript microRNA cDNA Synthesis Kit was used in an RT-qPCR reaction to amplify the human miR-1246 and miR-21 from a dilution series of HeLa total RNA isolated using Norgen’s Total RNA Purification Kit. The PCR target was robustly amplified from a broad range of RNA inputs, from the 200 ng down to 2 pg. Image Credit: Norgen Biotek Corp.
Figure 2. Robust Reverse Transcription of All RNA Transcripts (microRNAs and mRNA). Norgen’s microScript microRNA cDNA Synthesis Kit could effectively reverse transcribe both microRNA and other RNA species such as mRNA and large RNAs, allowing the parallel study of microRNA and its targets. HeLa RNA (200 ng) was used as a template in a reverse transcription using the microScript microRNA cDNA Synthesis Kit. The resulting cDNA was used in three different qPCR reactions for three different species of RNA. The top three panels showed the successful PCR amplification curve for miR-21 (microRNA), 5S rRNA (RNA without poly-A tail) and S15 (large mRNA). The lower three panels showed that each of the three PCR amplification showed unique amplicon with unique melt curve characteristics. Image Credit: Norgen Biotek Corp.
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Storage conditions and product stability
For a year following the date of dispatch, this kit remains stable.
Source: Norgen Biotek Corp.
| Component |
Cat. 54415 (12 rxns) |
Cat. 54410 (50 rxns) |
| microScript microRNA Enzyme Mix |
12 μL |
50 μL |
| 2x Reaction Mix |
120 μL |
500 μL |
| Universal PCR Reverse Primer |
60 μL |
250 μL |
| Nuclease-Free Water |
1.25 mL |
2 x 1.25 mL |
| Product Insert |
1 |
1 |