SuperScript™ IV CellsDirect™ cDNA Synthesis Kit
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SuperScript™ IV CellsDirect™ cDNA Synthesis Kit
Invitrogen™

SuperScript™ IV CellsDirect™ cDNA Synthesis Kit

Green features
The SuperScript IV CellsDirect cDNA Synthesis Kit is optimized for the synthesis of first-strand cDNA directly from a mammalian cellRead more
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Catalog NumberNo. of Reactions
1175015050 Reactions
11750350500 Reactions
Catalog number 11750150
Price (USD)
794.00
Each
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No. of Reactions:
50 Reactions
Request bulk or custom format
Price (USD)
794.00
Each
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The SuperScript IV CellsDirect cDNA Synthesis Kit is optimized for the synthesis of first-strand cDNA directly from a mammalian cell lysate without first isolating RNA. Cell lysis and reverse transcription are performed in the same tube, and the resulting first-strand cDNA is ready to use in PCR and qPCR.

Features of the SuperScript IV CellsDirect Synthesis Kit
• Compatible with a wide range of mammalian cell types grown under different treatment conditions
• Single-tube format minimizes reagent loss, sample loss, and handling time
• Total lysate volume can be used in first-strand cDNA synthesis reaction, providing greater yields with a limited number of cells and allowing for detection of rare transcripts
• High-quality cDNA generated for use in a variety of applications, including PCR and qPCR
• Simple protocol takes approximately 37 minutes
• Compatible with TaqMan and SYBR Green qPCR master mixes from various manufacturers
• Optimized performance for 1–10,000 cells per sample, results equivalent to those from purified RNA

Features of the SuperScript IV Reverse Transcriptase included in this kit
• Reduced RNase H activity
• High thermal stability
• High yields of cDNA in the first-strand synthesis reaction for greater sensitivity and enhanced detection of rare transcripts
• Superior tolerance to various RT inhibitors

SuperScript IV reverse transcriptase is a top choice for all RT-PCR and qRT-PCR applications. It is a proprietary MMLV mutant with superior robustness and reliability in RT reactions. It is significantly improved over SuperScript III reverse transcriptase in inhibitor resistance, processivity, and reaction speed, while retaining all the benefits of the previous enzyme, including increased thermostability, highly efficient full-length cDNA synthesis, and reduced RNase activity.

How it works
In traditional RT-PCR, RNA is first isolated from cells in a time-consuming procedure that can lead to a loss of material. Using the SuperScript IV CellsDirect cDNA Synthesis Kit, the cells are lysed and the cDNA is generated from the lysate in a single tube with minimal handling and no sample loss. DNase I treatment is an optional step and can be used to eliminate genomic DNA prior to first-strand synthesis. This kit has been optimized for small cell samples, ranging from 10,000 cells down to a single cell.

Specifications
FormatKit
GC-Rich PCR PerformanceHigh
Reaction Speed10 min.
TechniqueReverse Transcription
Green FeaturesLess Hazardous, Less Waste
Optimal Reaction Temperature50°C
Reverse TranscriptaseSuperScript IV
Ribonuclease H ActivityReduced
Shipping ConditionDry Ice
For Use With (Application)RT-PCR, Real Time PCR (qPCR)
Final Product TypeFirst-Strand cDNA
No. of Reactions50 Reactions
Reaction FormatPremixed Components
Reagent TypeReverse Transcription
Starting MaterialCells
Unit SizeEach
Contents & Storage

• SuperScript IV CellsDirect Lysis Solution (2 x 1.25 mL)
• SuperScript IV CellsDirect Stop Solution (1 x 250 μL)
• Lysis Enhancer (1 x 25 μL)
• Dnase I (1 x 55 μL)
• SuperScript IV RT Master Mix (1 x 400 μL)
• SuperScript IV No RT Control (1 x 400 μL)

Store at –20°C.

Frequently asked questions (FAQs)

What are the RT primers included in the SuperScript IV CellsDirect cDNA Synthesis Kit (Cat. No. 11750150, 11750350)?

The RT master mix contains both random and oligo dT primers.

What primers come in the SuperScript IV CellsDirect cDNA Synthesis Kit (Cat. No. 11750150)? Is it Oligo(dT) primers or random hexamers?

It is both. The RT master mix includes both, random hexamers and Oligo(dT) primers.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

In your SuperScript IV RT protocols, there is no ezDNase inactivation step. Will active ezDNase affect RNA or the RT reaction?

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.

Which SuperScript IV RT format do you recommend for real-time PCR applications?

For RT-qPCR applications we recommend using the Invitrogen SuperScript IV VILO Master Mix (Cat. No. 11756050). 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.

Are there any significant changes in the SuperScript IV RT protocol compared to the SuperScript III RT protocol?

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.