Invitrogen™ SuperScript™ III CellsDirect™ cDNA Synthesis Kit

Optimized to synthesize first-strand cDNA directly from a mammalian cell lysate without first isolating the RNA. Invitrogen™ SuperScript™ III CellsDirect™ cDNA Synthesis Kit includes the reagents, buffer, primer and other materials needed for lysis and reverse transcription (except DNA polymerase).

Catalog Number	Mfr. No.	No. of Reactions
18-080-300	Invitrogen™ 18080300	100 Reactions
18-080-200	Invitrogen™ 18080200	25 Reactions

Description

The SuperScript III CellsDirect cDNA Synthesis Kit is optimized for synthesizing first-strand cDNA directly from a mammalian cell lysate without first isolating the RNA. Lysis and reverse transcription are performed in the same tube, and the resulting first-strand cDNA is ready to use in cloning and PCR. For real-time quantitative RT-PCR, see the note below.

• 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 is used in first-strand cDNA synthesis reaction, providing greater yields with a limited number of cells and allowing for detection of rare transcripts
• SuperScript III Reverse Transcriptase, with reduced RNase H activity and higher thermal stability, produces high yields of cDNA in the first-strand synthesis reaction, for greater sensitivity and enhanced detection of rare transcripts
• Generates high-quality cDNA for use in a variety of applications, including cloning and PCR
• Simple protocol takes less than 2 hours

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 III CellsDirect cDNA Synthesis System, 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 is added 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 (as measured by serial dilution). The use of SuperScript III Reverse Transcriptase ensures high specificity and high yields of cDNA from small amounts of starting material—as little as 10pg total RNA. After synthesis, the first-strand cDNA can be amplified with specific primers by PCR without intermediate organic extractions or ethanol precipitations. Optimal reaction temperature: 50°C.

For real-time quantitative RT-PCR (qRT-PCR) from cell lysate, the Cells-to-CT products is recommened for RNA extraction. These kits include reagents and protocols that have been specifically optimized for real-time qRT-PCR.

PCR & Real-Time PCR, Reverse Transcription, cDNA Synthesis Direct from Cells

Specifications

• Detection Method: Primer-Probe
• Format: cDNA Synthesis Kit
• PCR Method: RT-PCR, RT-qPCR
• Reaction Speed: 50 min.
• Size (Final Product): Up to 4.5 kb
• Technique: Reverse Transcription
• Reverse Transcriptase: SuperScript III
• For Use With (Application): Real-Time Quantitative PCR (qPCR), Reverse Transcriptase PCR (RT-PCR)

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

For Research Use Only. Not for use in diagnostic procedures.