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Invitrogen™ SuperScript™ III CellsDirect™ cDNA Synthesis Kit DFS Item

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).

Supplier:  Invitrogen™ 18080300

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Catalog No. 18080300



Description

Description

The SuperScript™ III CellsDirect™ cDNA Synthesis Kit utilizes a simple protocol that takes less than 2 hours. Lysis and reverse transcription are performed in the same tube, and the resulting first-strand cDNA is ready to use in cloning and PCR.

  • 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

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.

Specifications

Specifications

• Resuspension Buffer, 1 mL
• RNaseOUT™ Recombinant Ribonuclease Inhibitor, 200 μL (40 units/μL)
• DNase I, 500 μL (1 U/μL)
• 10X DNase I Buffer, 160 μL
• 25 mM EDTA, 120 μL
• Oligo(dT)20, 120 μL (50 μM)
• 10 mM dNTP Mix, 100 μL
• SuperScript™ III RT, 100 μL (200 units/μL)
• 5X RT Buffer, 600 μL
• DTT, 100 μL (0.1 M)
E. coli RNase H, 100 μL (2 U/μL)
• HeLa Total RNA, 10 μL (10 ng/μL)
• Forward/Reverse Control Primers, 10 μL (10 μM)
E. coli RNase H (2 U/μL)

Store all components at –20°C.

First-Strand cDNA
High
RT-PCR, RT-qPCR
50 min.
Up to 4.5 kb
Reverse Transcription
SuperScript III
Real-Time Quantitative PCR (qPCR), Reverse Transcriptase PCR (RT-PCR)
Primer-Probe
cDNA Synthesis Kit
100 Reactions
Separate components
Reverse Transcription
Cells
50°C
Reduced
SDS
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For Research Use Only. Not for use in diagnostic procedures.