Invitrogen™ SuperScript™ IV First-Strand Synthesis System with ezDNase™ Enzyme

SuperScript™ IV First-Strand Synthesis System for RT-PCR is optimized for synthesis of first-strand cDNA from purified poly(A)+ or total RNA. Reliable, consistent, and fast cDNA synthesis even in the presence of inhibotors, with all components needed for gDNA removal and RT reaction.

Catalog Number	Mfr. No.	No. of Reactions
18-091-150	Invitrogen™ 18091150	50 Reactions
18-091-300	Invitrogen™ 18091300	200 Reactions

Description

• Kit includes SuperScript IV Reverse Transcriptase (RT), a proprietary MMLV mutant that produces robust and reliable cDNA synthesis in RT reactions, and ezDNase to remove potential genomic DNA contamination
• System is significantly improved over the SuperScript III-based system in inhibitor resistance, processivity, and reaction speed, while retaining all the benefits of the previous system, including increased thermostability, highly efficient full-length cDNA synthesis, and reduced RNase activity
• 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
• Contains all components needed for RT reactions, plus an additional control gene and primers, and provides flexibility to customize the RT set-up
• ezDNase for genomic DNA removal is included
• Simplified genomic DNA removal step dramatically reduces the time of the entire reverse transcription protocol and reduces possible variation in gene expression due to RNA loss or damage during conventional DNase treatment
• Ideal for when reproducibility and reliability is the primary concern and when inhibitors in the RNA sample may interfere with cDNA synthesis, leading to biases in gene expression studies
• Improved resistance to a variety of inhibitors that can interfere with cDNA synthesis
• Robust and specific cDNA synthesis in a wide range of sample types
• Faster reverse transcriptase reaction that reduces incubation time from >50 min to 15 min including a simplified genomic DNA removal step

 

Enzyme 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.

Performance and quality testing
Endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product.

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

Specifications

• Detection Method: Primer-Probe
• Format: First-Strand Synthesis System
• PCR Method: RT-PCR, RT-qPCR
• Reaction Speed: 10 min.
• Size (Final Product): Up to 12.3 kb
• Technique: Reverse Transcription
• Reverse Transcriptase: SuperScript IV

• Final Product Type: First-Strand cDNA
• GC-Rich PCR Performance: High
• Reaction Format: Separate components
• Reagent Type: Reverse Transcription
• Starting Material: RNA
• Optimal Reaction Temperature: 50°C