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.
For Laboratory Use
For Use With (Application):
RT-PCR (Endpoint), Real-Time PCR
SuperMix or Master Mix
Contents & storage
• 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.