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SuperScript IV CellsDirect includes a powerful SuperScript IV reverse transcriptase enzyme and a simplified cDNA synthesis workflow that eliminates the need to purify RNA. With this basic approach in a single tube, you can avoid RNA loss and contamination by reducing the number of pipetting steps and manipulations. The end result is saving time and effort.







 

Superior Sensitivity and Yield 

The SuperScript IV CellsDirect cDNA synthesis kit consistently outperforms kits from competitors. It is sensitive across a range of cell numbers and sample sizes and produces greater yields for abundant (ACTB), moderately abundant (PGK1), and low abundance (BCL2) transcripts in studies with Hela S3 cells.

High Abundance Cell Target - ACTB

The SuperScript IV CellsDirect cDNA synthesis kit was used to synthesize cDNA from ACTB, an abundant transcript, using target specific primers. Figures illustrate the superior sensitivity and yield compared to the kits of competitors.
 

ACTB cDNA synthesis analyzed with a SYBR green assay

cDNA synthesis of ACTB analyzed using a SYBR Green assay. Using a serial dilution of HeLa S3 cells representing a range from 10 to 10,000 cells per sample, cells were lysed and cDNA synthesized according to each kit manufacturer’s specifications. Analysis was performed using the PowerTrack SYBR Green Master Mix.	Bar graph showing a comparison of the CT difference, which represents the yield. Graph uses the same data from the results presented above demonstrating sensitivity and yield for high abundance transcripts (ACTB).

 

Sensitivity and Yield - Low Abundance Gene Target - BCL2

To demonstrate sensitivity and yield for low abundance targets, the SuperScript IV CellsDirect cDNA synthesis kit was used to synthesize cDNA from BCL2 using target specific primers. Figures illustrate the superior sensitivity and yield across the full 10-fold dilution series of HeLa S3 cells (10 to 10,000 cells) as compared to the kits of competitors.
 

BCL2 cDNA synthesis analyzed with a SYBR green assay
 

cDNA synthesis of BCL2 and analyzed using a SYBR Green assay. Using a serial dilution of HeLa S3 cells representing a range from 10 to 10,000 cells per sample, cells were lysed and cDNA synthesized according to each kit manufacturer’s specifications. Analysis was performed using the PowerTrack SYBR Green Master Mix.	Bar graph using the same data as above showing the product vs. competitor kits across dilution series demonstrating sensitivity and yield for low abundance transcripts (BCL2).

 

Sensitivity and Yield - Moderate Abundance Gene Target - PGK1

To further demonstrate sensitivity and yield, SuperScript IV CellsDirect cDNA synthesis kit was used to synthesize cDNA from PGK1, a moderately abundant gene target, using target specific primers. Figures illustrate sensitivity and yield across a 10-fold dilution series of HeLa S3 cells (10 to 10,000 cells) as compared to the kits of competitors.
 

cDNA synthesis of PGK1 and analysis using a TaqMan assay. Using a serial 10-fold dilution of HeLa S3 cells representing a sample sizes from 10 to 10,000 cells per sample, cells were lysed and cDNA synthesized according to each kit manufacturer’s specifications. Analysis was performed using the TaqMan Fast Advanced Master Mix.	Bar graph uses the same data as above showing the product vs competitor kits across dilution series demonstrating sensitivity and yield for moderately abundant transcripts (PGK1).

Sensitivity and Yield - iPSC

The SuperScript IV CellsDirect cDNA synthesis kit consistently outperformed competitors for a range of low to high abundant genes across many cell lines including Induced Pluripotent Stem Cells (iPSC).
 

Higher reverse transcriptase efficiency with SuperScript IV CellsDirect cDNA synthesis Kit than with other commercial kits (SuperScript III CellsDirect cDNA synthesis Kit (Invitrogen), SingleShot Probes Kit (Bio-Rad), FastLane Cell cDNA Kit (Qiagen),  RealTime ready Cell Lysis Kit + Transcriptor First Strand cDNA Synthesis Kit (Roche). cDNA synthesis was performed from 1000 human iPSC stem cells inputs to lysis and analyzed with various from low to high expression gene primers in TaqMan assay.

 

Linearity

The SuperScript IV CellsDirect cDNA synthesis kit consistently outperformed competitors during linearity tests.

SuperScript™ IV CellsDirect™ cDNA Synthesis Kit

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 kit include:

• 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 around 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

The SuperScript IV CellsDirect cDNA Synthesis Kit includes SuperScript IV Reverse Transcriptase. Features of this enzyme include:

• Reduced RNase H activity
• Higher 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.

Content&Storage

SuperScript™ IV CellsDirect™ cDNA Synthesis Kit / 50 Reaction

• SuperScript IV CellsDirect Lysis Solution, 1.25 mL
• SuperScript IV CellsDirect Stop Solution, 250 µL
• Lysis Enhancer, 25 µL
• Dnase I, 55 µL
• SuperScript IV RT Master Mix, 400 µL
• SuperScript IV No RT Control, 400 µL

SSIV CellsDirect Lysis Solution can be stored it at 4°C. The other kit components (Stop Solution, DNase I, Lysis Enhancer, SSIV RT Master Mix, and SSIV No RT Control) should be stored at –20°C.

SuperScript™ IV CellsDirect™ cDNA Synthesis Kit / 500 Reaction

• SuperScript IV CellsDirect Lysis Solution, 9 mL
• SuperScript IV CellsDirect Stop Solution, 1.25 mL
• Lysis Enhancer, 250 µL
• DNase I, 500 µL
• SuperScript IV RT Master Mix, 1 mL
• SuperScript IV No RT Control, 1 mL

SSIV CellsDirect Lysis Solution can be stored it at 4°C. The other kit components (Stop Solution, DNase I, Lysis Enhancer, SSIV RT Master Mix, and SSIV No RT Control) should be stored at –20°C.
 

SuperScript™ IV CellsDirect™ cDNA Synthesis Kit / 50 Reaction
Sample Type	Cells
Shipping Conditions	Dry Ice
Detection Method	Fluorescence
For use with Equipments	RT-PCR (Endpoint), Real-Time PCR
Format	Tube
GC-Rich PCR Performance	High
Number of Reaction	50 Reaction
Number of Test	50 Reaction
PCR Method	qPCR, RT-PCR
Product Line	SuperScript™
Reaction Format	SuperMix or Master Mix

 

SuperScript™ IV CellsDirect™ cDNA Synthesis Kit / 500 Reaction
Sample Type	Cells
Shipping Conditions	Dry Ice
Detection Method	Fluorescence
For use with Equipments	RT-PCR (Endpoint), Real-Time PCR
Format	Tube
GC-Rich PCR Performance	High
Number of Reaction	500 Reaction
Number of Test	500 Reaction
PCR Method	qPCR, RT-PCR
Product Line	SuperScript™
Reaction Format	SuperMix or Master Mix

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