Description
MicroRNAs are a class of non-coding RNAs with a length of about 22 nt, which play an important role in the regulation of gene expression in plants and animals. This kit uses Poly(A)-tailing method to perform reverse transcription from miRNA first-strand to cDNA. The 2×Hifair™ miRNA RT buffer in the product contains all the raw materials and primers for miRNA Poly(A)-tailing reaction and reverse transcription reaction. Careful optimization ensures that Poly(A) modification process and reverse transcription process can be performed simultaneously and efficiently in miRNA 3' end.
This product is recommended to be used in conjunction with our company's Hieff™ miRNA Universal qPCR SYBR Master Mix (Cat#11171) for optimal experimental results.
Features
- High sensitivity detection rate: 10 pg RNA can be detected
- High product stability: remains unaffected at 37℃ for 7 days
Application
- First-strand cDNA is synthesized and can be used as a template for RT-PCR and real-time RT-qPCR.
Specifications
| Final product type | cDNA (First-Strand) |
| PCR method | RT-PCR |
| Reaction format | Isolated fraction |
| Sample type | miRNA |
| Optimal reaction temperature | 37℃ |
| Reverse transcriptase | MMLV |
Components
| Components No. | Name | 11148ES50 (50T) |
| 11148-A | Hifair™ miRNA RT enzyme mix | 87.5 μL |
| 11148-B | 2×Hifair™ miRNA RT buffer | 250 μL |
| 11148-C | RNase-free H2O | 10 mL |
| 11148-D | Universal Reverse Primer (10 μmol/L) | 4 mL |
| 11148-E | U6 Forward Primer (10 μmol/L) | 500 μL |
| 11148-F | U6 Reverse Primer (10 μmol/L) | 500 μL |
Storage
The product should be stored at -25°C~-15℃ for 1 year.
Figures
Figure1. High sensitivity detection rate: 10 pg RNA could be detected.
The synthesized hsa-miR-let-7e-5p (a) and 293T cell total RNA (b) were used as templates and diluted to the following gradients: 60 copies to 606 copies (6 gradients) and 10 pg-100 ng (5 gradients). All gradient cDNA was detected.
[1] Huang G, Chen J, Liu J, Zhang X, Duan H, Fang Q. MiR-935/HIF1α Feedback Loop Inhibits the Proliferation and Invasiveness of Glioma. Onco Targets Ther. 2020;13:10817-10828. Published 2020 Oct 23. doi:10.2147/OTT.S244409(IF:4.147)
[2] Li H, Pan R, Lu Q, et al. MicroRNA‑145‑5p inhibits osteosarcoma cell proliferation by targeting E2F transcription factor 3. Int J Mol Med. 2020;45(5):1317-1326. doi:10.3892/ijmm.2020.4504(IF:4.101)