OneScript® Hot 5X RT MasterMix

Cat. No.

G590

Unit

100 rxn (400 µl)

Price

$209.00 $189.00

Cat. No.

G590

Name

OneScript® Hot 5X RT MasterMix

Unit

100 rxn (400 µl)

Category

Reverse Transcriptase & RT-PCR

Description

OneScript® Hot 5X RT MasterMix is a convenient, ready-to-use formulation designed for efficient first-strand cDNA synthesis. This product simplifies the cDNA synthesis process, ensuring reliability and reproducibility. The optimized 5X RT MasterMix contains abm’s proprietary OneScript® Hot Reverse Transcriptase, RNaseOFF Ribonuclease Inhibitor, dNTPs, and a carefully balanced ratio of Oligo(dT)s and Random Primers. The resulting high-quality cDNA is suitable for a wide range of downstream applications.

Product Features:

Easy one-step setup reduces pipetting and sample handling
High cDNA yields from difficult samples with OneScript^® Hot Reverse Transcriptase (Cat. No. G593)
Inhibits ribonuclease contaminants with RNaseOFF Ribonuclease Inhibitor (Cat. No. G591)
Also includes dNTPs, Oligo (dT)s, and random primers
Included in abm's PCR Buffet Program

Product Component	Quantity
OneScript® Hot 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

If input RNA samples are expected to have high levels of genomic DNA contamination, we recommend using abm’s All-In-One 5X RT MasterMix with gDNA Removal (Cat. No. G592).

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. G590

Datasheet

OneScript® Hot 5X RT MasterMix Datasheet

Search CoA here

MSDS

OneScript® Hot 5X RT MasterMix MSDS

Other

	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.

	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 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.

	What if input RNA samples are expected to have high levels of genomic DNA contamination?
	If input RNA samples are expected to have high levels of genomic DNA contamination, we recommend using abm’s All-In-One 5X RT MasterMix with gDNA Removal (Cat. No. G592).

What is the advantage of OneScript® Hot Reverse Transcriptase compared to OneScript® Plus Reverse Transcriptase?

OneScript® Hot Reverse Transcriptase offers a significant advantage over enzymes that operate at lower temperatures (42-55ºC) due to its exceptional thermostability, allowing it to function effectively at 60-72ºC. This higher temperature improves efficiency by preventing RNA secondary structures and enabling cDNA synthesis from degraded or low-abundance RNA. Additionally, it is more resistant to inhibitors and contaminants, making it ideal for complex or impure RNA samples.

	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. G590 equivalent to?
	This product is abm’s next generation of RT MasterMixes and is functionally equivalent to G451, G452, G453, G454 (OneScript® cDNA Synthesis SuperMixes), with improved performance. Contact our customer service team technical@abmgood.com for more information.

	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.