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All-In-One 5X RT MasterMix with gDNA Removal

Cat. No.

G592

Unit

100 rxn

Price

$209.00

Cat. No.

G592

Name

All-In-One 5X RT MasterMix with gDNA Removal

Unit

100 rxn

Category

Reverse Transcriptase & RT-PCR

Description

The All-In-One 5X RT MasterMix with gDNA Removal offers a convenient, ready-to-use solution for first-strand cDNA synthesis with genomic DNA removal, all in a single tube. This powerful formulation addresses the common challenge of genomic DNA contamination, ensuring accurate RNA detection without compromising reverse transcription or cDNA synthesis.

The optimized MasterMix includes abm’s proprietary OneScript® Hot Reverse Transcriptase, RNaseOFF Ribonuclease Inhibitor, temperature-sensitive DNase, dNTPs, and a carefully balanced mix of Oligo (dT)s and Random Primers. By eliminating the need for separate reagents, it provides a straightforward, reliable, and reproducible cDNA synthesis process with high yields—even from difficult samples. The high-quality cDNA produced is suitable for a wide range of downstream applications.

Product Features:

Easy one-step setup reduces pipetting and sample handling
Removes contaminating gDNA with temperature-sensitive DNase
High cDNA yields from difficult samples with OneScript^® Hot Reverse Transcriptase (Cat. No. G593)
Inhibits ribonuclease contaminants with RNaseOFF Ribonuclease Inhibitor (Cat. No. G138)
Also includes dNTPs, Oligo (dT)s, and random primers
Included in abm's PCR Buffet Program

Product Component	Quantity
All-In-One 5X RT MasterMix	100 rxn (400 µl)
Nuclease-Free H₂O	1.0 ml

ISO 13485:2016 MDSAP Certified
Our PCR Products are manufactured under a Quality Management System conforming with ISO 13485:2016 as certified by Intertek (a MDSAP recognized auditing organization).

Application

Generation of templates for use in RT-PCR and qRT-PCR
cDNA synthesis from ssRNA
cDNA library construction
Generation of probes for hybridization
DNA primer extension

Concentration

Storage Condition

Store at -20°C.

Note

For applications using pure RNA where genomic DNA removal is not required, try our OneScript® Hot 5X RT MasterMix (Cat. No. G590). It offers a faster protocol for an even more efficient cDNA synthesis.

Material Citation

If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G592

Datasheet

G592 Datasheet

Search CoA here

MSDS

G592 MSDS

Supporting Protocol

Other

PCR Product Guide

	What's included in this kit?
	The kit includes everything you need for cDNA synthesis in one tube: All-In-One 5X RT MasterMix (100 reactions, 400 µl), which contains reverse transcriptase, RNaseOFF RNase inhibitor, DNase, dNTPs, and primers. Additionally, a tube of Nuclease-Free H₂O (1.0 ml) is provided to complete your reaction setup.

	How should I store the All-In-One 5X RT MasterMix with gDNA Removal?
	Store the All-In-One 5X RT MasterMix with gDNA Removal at -20°C. This product is unstable if stored improperly. Reduce freeze-thaw cycling and handle with care.

	Can I use both total RNA and poly(A)+ mRNA for cDNA synthesis?
	Yes, both total RNA and poly(A)+ mRNA can be used, though poly(A)+ mRNA typically yields higher quantities and better purity.

	How can I enhance genomic DNA removal?
	To improve genomic DNA removal, increase the 37°C incubation time from 15 minutes to 30 minutes.

	Can I synthesize cDNA from long RNA transcripts?
	Yes, for longer RNA transcripts, extend the 60°C incubation time to up to 30 minutes.

	What if genomic DNA removal isn’t necessary?
	If genomic DNA removal is not required, use the OneScript® Hot 5X RT MasterMix (Cat. No. G590) for a faster and more efficient cDNA synthesis process.

	What can I use the synthesized cDNA for?
	The high-quality cDNA can be used in a variety of downstream applications, including gene expression analysis, cloning, and PCR-based assays. BlasTaq™ 2X qPCR Master Mix (Cat. No. G891) is well suited to downstream qPCR applications.

	How should I store the synthesized cDNA?
	Store the synthesized first-strand cDNA at -20°C for long-term use.

Why am I getting low cDNA yields?

Low cDNA yields can be caused by poor RNA integrity, contamination, or insufficient RNA input. To improve yield:

Check RNA integrity with gel electrophoresis or a BioAnalyzer, aiming for a minimum A260/A280 ratio of 1.7 or a RIN above 8.
Clean the RNA using ethanol precipitation or lithium chloride to remove contaminants.
Purify the RNA with extraction kits or phenol/chloroform to remove proteins.
Increase RNA input, especially for low-abundance samples.

	How much RNA template should I use for cDNA synthesis?
	We recommend using 1 ng to 2 μg of RNA per reaction.

	What volumes of cDNA should I use for downstream PCR?
	Typically, use 1 μl of cDNA in a 25 μl PCR reaction. You can add up to 20% of the PCR volume (e.g., 5 μl in a 25 μl PCR), depending on your target and primers.

	Do I need an RNase inhibitor when using this product?
	All of abm's Reverse Transcriptases include RNaseOFF Ribonuclease Inhibitor, which protects RNA from degradation and is resistant to oxidation, even under low DTT concentrations. Therefore, there’s no need to add an external RNase inhibitor, as RNaseOFF provides optimal RNA protection during reverse transcription.

What discontinued abm products is Cat. No. G592 equivalent to?

This product is abm's next generation of RT MasterMixes and is functionally equivalent to Cat. No. G485, G486, G490, G492, G916 (5X All-In-One RT MasterMixes with AccuRT Genomic DNA Removal Kit or with ExCellenCT Lysis Kits), and G451, G452, G453, G454 (OneScript® cDNA Synthesis SuperMixes), with improved performance. For more information, contact our customer service team at technical@abmgood.com.

	What is the source of this enzyme?
	The enzyme is produced recombinantly in E. coli, which has been engineered to express the enzyme gene. While the original gene may come from another organism, all production and purification occur using E. coli under controlled conditions.

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