To help life science research,
let more research results benefit mankind
To help life science research,
let more research results benefit mankind
1) There are DNase and RNase inhibitors in the master mix.
2) Genome removal and reverse transcription can be completed in one step.
3) The reaction temperature was 50°C.
Highlights
1) Simple and fast: genome removal and reverse transcription can be completed in one or two steps. Just add RNA template and water to perform reverse transcription reaction;
2) Strong thermal stability: the reaction at 50℃ improves the reverse transcription efficiency of the complex secondary structure of the RNA template;
3) High reverse transcription efficiency: Compared with similar products on the market, the reverse transcription efficiency of this product is higher.
Introduction
This product is an efficient and convenient first-strand cDNA synthesis reagent. The UnionScript First-strand cDNA Synthesis Mix contains UnionScript Reverse Transcriptase and its reaction Buffer, Rnasin, dNTPs, Oligo(dT)20VN and random primers and other first-strand cDNAs all the components required for synthesis are ready for reverse transcription by adding RNA template and water.
The UnionScript Reverse Transcriptase contained in this product is a new generation of reverse transcriptase, 50℃ reaction, strong thermal stability, high reverse transcription efficiency. dsDNase can effectively remove the residual genomic DNA in the RNA template, ensuring more reliable subsequent quantitative results, and qPCR primers do not need to be designed across introns. The cDNA obtained by reverse transcription of this product can be used for downstream qPCR detection.
Composition
Contents | Amount(100 rxns) |
UnionScript First-strand cDNA Synthesis Mix | 400 μL |
dsDNase | 2×50 μL |
10×dsDNase Buffer | 200 μL |
Nuclease-Free Water | 2×1.0 mL |
Related products
Type | Name | Cat. No. | Amount | Introduction |
RNA Extraction | RE704 | 50 rxns | This product is suitable for total RNA extraction from plant samples. | |
RNA Extraction | RE705 | 50 rxns | Unique genomic DNA filter column without DNase treatment; No toxic reagents such as phenol, chloroform and β-mercaptoethanol are required. | |
qPCR | SQ412 | 100 rxns | Using antibody-modified new hot-start GS AntiQ DNA polymerase and carefully optimized Buffer. It is equipped with two ROX Reference Dye (High/Low) according to different instrument models. | |
High-Fidelity PCR | SF212 | 5×1.125 mL | It is a 1.1 × ready-to-use mixture for efficient PCR amplification, which has high fidelity and extremely fast extension speed. | |
Conventional PCR | ST111 | 5×1.0 mL | It is a basic Taq DNA polymerase with wide applicability, high yield and extremely fast extension speed. |
Q1：Why is the CT value high after qPCR detection of reverse transcribed cDNA products?
A1：
1) Template RNA extraction failed or degraded. It is recommended to verify the integrity of RNA by electrophoresis before reverse transcription reaction, and perform reverse transcription and qPCR operations as soon as possible after RNA extraction;
2) The gene expression level is low. The amount of template RNA can be appropriately increased;
3) Improper operation of reverse transcription system, program or sample addition. It is recommended to operate according to the specifications of the manual.
Q2：Why is the melting curve not a single peak after performing qPCR detection after reverse transcription?
A2：
1) RNA template contains gDNA. It is recommended to perform degenome reaction or primer cross-intron design before reverse transcription.
2) Poor specificity of qPCR primers. Redesign primers with better specificity, and the specificity of primers can be detected by conventional PCR.
3) Excessive primers. Excessive primers may form primer-dimers, and it is recommended to add them according to the amount recommended in the instructions.
4) The system may be polluted. Set up a no-template negative control (NTC) to check whether the system is contaminated. If there is contamination, it is recommended to replace the reagents and consumables, and use a nucleic acid cleaner (Cat. No. : ND709) to clean the work surface and aerosol contamination in the experimental environment.
Manual: Click to download
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