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RT Supermix for qPCR

Reverse Transcription Supermix for qPCR

Catalog No. Product Name Size List Price (US$) Quantity
MB0127-5G 5G RT Supermix for qPCR with gDNA Elimination Function 100 reactions (20 ul/reaction) 280.00
MB0127-4G 4G RT Supermix for qPCR with gDNA Elimination Function 100 reactions (20 ul/reaction) 220.00
Description

MB0127-5G: 5G RT Supermix for qPCR with gDNA Elimination Function

The 5G RT supermix for qPCR with gDNA elimination function provides a fast and efficient procedure for reverse transcription and genomic DNA elimination in RNA samples. To obtain accurate results in RT-qPCR, it is important that only cDNA is amplified and detected. Interference by genomic DNA can be avoided by designing primers or probes that span an exon/exon boundary. However, in cases where this is not possible (e.g., the cDNA is from a single-exon gene, or there exist processed pseudogenes in genomic DNA sequences), it is essential that the starting RNA sample is free of genomic DNA. The 5G RT supermix for qPCR with gDNA elimination function combines genomic DNA elimination of RNA samples and first strand cDNA synthesis in one easy workflow to ensure accuracy of RT-qPCR results. Since there is no need to perform a separate DNase digestion and re-purify your RNA samples, your time and costs are saved.

The 5G RT supermix for qPCR with gDNA elimination function is suitable for two-step RT-qPCR detection, and compatible with dye-based and probe-based qPCR, enabling high-performance gene expression analysis. Genomic DNA elimination and reverse transcription can be simultaneously completed, which is convenient, and can greatly reduce contamination and RNA degradation. The new generation of reverse transcriptase, 5G Reverse Transcriptase, is the most suitable for reverse transcription optimization. The heat-labile DNase is highly efficient, and can be easily inactivated. Buffer was optimized to improve the efficiency of one-strand synthesis. The 5G RT supermix contains single-component reverse transcription primers Oligo (dT)20 VN and Random hexamers. Users can choose reverse transcription primers for subsequent experiments according to your needs. The 5G RT supermix can be used to synthesize highly uniform cDNA for qPCR quantification.

Features of the 5G RT supermix for qPCR with gDNA elimination function:

* Simple and fast operation: one-step genome clearance and reverse transcription.
* Higher reverse transcription efficiency and higher cDNA yields from all regions of RNA transcripts: based on the more efficient 5G Reverse Transcriptase.
* Flexible choice of primers: Different types of reverse transcription primers can be used flexibly for different experimental designs.
* Excellent cDNA stability: complete deactivation of thermal DNase and long-term storage of cDNA.

Components of the 5G RT supermix for qPCR with gDNA elimination function:

gDNA Removing Buffer (4X): Buffer for effective elimination of genomic DNA contamination in RNA samples
5G Enzyme Mix (5X): Buffer optimized for reverse transcription; Mixtures of MgCl2, dNTPs, stabilizers, 5G reverse transcriptase, RNase inhibitor, Oligo (dT)20 VN Primer, and Random Hexamers.
RT Negative Control Mix (5X): The same as 5G Enzyme Mix (5X) except 5G reverse transcriptase.
RNase-free H2O: Ultrapure quality.

Storage:

The kit is stable for one year when stored in a constant temperature freezer at -20°C. After thawing, mix the components thoroughly before using. Frequent freezing and thawing is not recommended.

MB127-4G: 4G RT Supermix for qPCR with gDNA Elimination Function

The 4G RT supermix for qPCR with gDNA elimination function is suitable for two-step RT-qPCR detection, and compatible with dye-based and probe-based qPCR, enabling high-performance gene expression analysis. The 4G RT supermix for qPCR includes 4G Reverse Transcriptase, which is a new reverse transcriptase obtained through in vitro molecular evolution technology on the basis of M-MLV (RNase H-) Reverse Transcriptase. Compared with the previous generation of Reverse Transcriptase, the 4G Reverse Transcriptase has further greatly improved thermal stability and is very suitable for reverse transcription of RNA templates with complex secondary structures. With multiple point mutations which further enhances template affinity and progress, the 4G Reverse Transcriptase has higher tolerance to common reverse transcription inhibitors, which greatly improves the ability to synthesize full-length cDNA.

The 4G RT supermix for qPCR with gDNA elimination function contains all the components needed to synthesize high-quality first-strand cDNA. The 4 × gDNA Removing Buffer in this kit can quickly remove genomic DNA contamination at 42°C for 2 min, ensuring more reliable follow-up results, and simplifying qPCR primer design without the need to design primers across introns. The 4G RT supermix contains single-component reverse transcription primers Oligo (dT)20 VN and Random hexamers. Users can choose reverse transcription primers for subsequent experiments according to your needs. The 4G RT supermix can be used to synthesize highly uniform cDNA for qPCR quantification.

Features of the 4G RT Supermix for qPCR with gDNA elimination function:

* Based on the highly efficient 4G Reverse Transcriptase, the super temperature tolerance ensures that the complex secondary structure of RNA can be opened under high temperature conditions to obtain longer cDNA.
* Quick and complete removal of genome contamination by the 4 × gDNA Removing Buffer: to ensure more reliable follow-up results, and simplify the design of qPCR primers, without the need to design primers across introns.
* Different primers are selected for different downstream applications.

Components of the 4G RT Supermix for qPCR with gDNA elimination function:

gDNA Removing Buffer (4X): Buffer for effective elimination of genomic DNA contamination in RNA samples
4G Enzyme Mix (5X): Buffer optimized for reverse transcription; Mixtures of MgCl2, dNTPs, stabilizers, 4G reverse transcriptase, RNase inhibitor, Oligo (dT)20 VN Primer, and Random Hexamers.
RT Negative Control Mix (5X): The same as 4G Enzyme Mix (5X) except 4G reverse transcriptase.
RNase-free H2O: Ultrapure quality.

Guidelines for Reverse Transcription:

* High-quality RNA is essential for high-quality cDNA synthesis.
* Set up all reactions on ice to minimize the risk of RNA degradation.
* After reverse transcription, the reaction must be inactivated by incubation at 95°C for 3 minutes.
* Primers are supplied in this supermixes. If gene-specific primers (not supplied) should be used, we recommend using a final concentration of 0.7 uM, and a primer titration from 0.5 uM to 1 uM can be performed.
* For two-step RT-qPCR, the volume of the cDNAs added (from the undiluted RT reaction) should not exceed 1/10 of the final PCR volume.

Quality Control:

All components in the 4G and 5G RT supermixes for qPCR are free of contaminating DNase and RNase. The kits are functionally tested using real-time RT-PCR with SYBR Green qPCR Master Mix for amplification of GAPDH cDNA derived from human universal RNA.

PCR enzymes:
* Taq DNA polymerase.
* High yield Taq DNA polymerase.
* Hot-start high-fidelity DNA polymerase.
* Hot start Taq DNA polymerase with chemical or antibody inhibitor, used for SYBR green or Taqman probe-based real time PCR (qPCR or quantitative PCR).

2x PCR master mixes:
* 2x PCR PreMix with dye (green, red, blue).
* 2x rapid PCR Master Mix with a green dye.
* 2x long PCR Master Mix with a blue dye.
* 2x high yield PCR master mix with a blue dye.
* 2x hot-start high-fidelity PCR master mix with a blue dye.
* Blood direct PCR kit
* Cell and tissue direct PCR kit
* RT-PCR master mixes
* 2x qPCR master mixes
* RT-qPCR master mixes

PCR components and related products:
* dNTP mix, 10 mM each.
* dNTP mix, 25 mM each.
* Proteinase K powder .
* Proteinase K solution.
* Reverse Transcriptases.
* First strand cDNA synthesis kits.
* Murine RNase inhibitor.
* PCR enhancer.

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