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Catalog Number | No. of Reactions |
---|---|
18090010 | 10 Reactions |
18090050 | 50 Reactions |
18090200 | 200 Reactions |
SuperScript IV Reverse Transcriptase (RT) is a proprietary MMLV mutant with superior robustness and reliability in RT reactions. It is significantly improved over SuperScript III RT in inhibitor resistance, processivity, and reaction speed, while retaining all the benefits of that enzyme, including increased thermostability, highly efficient full-length cDNA synthesis, and reduced RNase activity. SuperScript IV RT is designed to provide reliable, consistent, and fast cDNA synthesis in the presence of inhibitors found in a wide variety of samples that cause other currently available RTs to perform inefficiently.
SuperScript IV RT offers advantages at standard as well as elevated temperatures over cloned AMV and ThermoScript RT, including higher sensitivity, inhibitor tolerance, and ability to work with degraded RNA.
Features of SuperScript IV Reverse Transcriptase include:
• Significantly improved resistance to inhibitors that can interfere with cDNA synthesis
• Robust and specific cDNA synthesis in a wide range of sample types
• Faster reverse transcriptase reaction reduces incubation time from >50 minutes to 10 minutes
• Better processivity when compared to SuperScript III RT
SuperScript IV RT is the top choice for all RT-PCR and qRT-PCR applications in the SuperScript product family when reproducibility and reliability is the primary concern. It is also our top performing RT when inhibitors in the RNA sample may interfere with cDNA synthesis, leading to biases in gene expression studies.
Source
Purified from E. coli expressing the pol gene of M-MLV, modified to increase thermostability and half-life, processivity, inhibitor resistance and to reduce RNase H activity
Unit definition
One unit of SuperScript IV RT is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min at 37°C using poly(A) oligo(dT)12-18 as a template/primer.
Unit reaction conditions
50 mM Tris-HCl (pH 8.3), 4 mM MgCl2, 10 mM DTT, 50 mM KCl, 0.5 mM dTTP, 0.4 MBq/mL [3H]-dTTP, 0.4 mM poly(A) oligo (dT)12-18 and enzyme in 20 μl for 10 min at 37°C.
• SuperScript IV RT (10 μL)
• 5X RT buffer (1 mL)
• 0.1 M DTT (500 μL)
Store at -15°C to -25°C.
Yes, you can use a DNA-RNA hybrid as a template for M-MLV Reverse Transcriptase.
We have not tested this for SuperScript reverse transcriptases, so we cannot guarantee it would also work with those products.
This article can be used as a reference for additional information.
Find additional tips, troubleshooting help, and resources within our Reverse Transcription and RACE Support Center.
The Invitrogen ezDNase Enzyme is a novel DNase that is highly specific for double-stranded DNA. It has no activity on single-stranded DNA in RT reactions (primers or probes), or on RNA. The enzyme is also thermolabile—it is inactivated quickly at temperatures typical for the SuperScript IV RT reaction (e.g., 50°C). The additional inactivation step is therefore not required in RT-qPCR applications.
Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.
For RT-qPCR applications we recommend using the Invitrogen SuperScript IV VILO Master Mix. The cDNA synthesis reaction setup with this master mix requires fewer pipetting steps and therefore reduces variation in the data. SuperScript IV RT, as a component of the master mix, offers the highest efficiency of cDNA synthesis step compared to competitors’ products.
Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.
The only change is that the incubation time for the reverse transcription reaction has been reduced from 50 minutes to 10 minutes. All the other parameters and steps are the same.
Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.
When compared with SuperScript III RT (and other manufacturers’ RTs) in a synthesis reaction for a 9 kb cDNA, SuperScript IV RT performed successful synthesis in just 10 minutes and did so with comparable (or improved) yield (as shown by gel band density).
Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.
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