Invitrogen™ SuperScript™ III CellsDirect™ cDNA Synthesis Kit

The SuperScript III CellsDirect cDNA Synthesis Kit is optimized for synthesis of first-strand cDNA directly from a mammalian cell lysate without first isolating the RNA.

Supplier:  Invitrogen™ 18080300

Catalog No. 18-080-300

Description

The SuperScript III CellsDirect cDNA Synthesis Kit is optimized for synthesis of 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.

Features of the SuperScript III CellsDirect Synthesis 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 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 two 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 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 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 10 pg total RNA). After synthesis, the first-strand cDNA can be amplified with specific primers by PCR without intermediate organic extractions or ethanol precipitations.

Specifications

• Content And Storage:

  • 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 at –20°C.
  
  

• GC-Rich PCR Performance: High
• Technique: Reverse Transcription
• Reverse Transcriptase: SuperScript III
• For Use With (Application): Real Time PCR (qPCR)
• No. of Reactions: 100 Reactions
• Reagent Type: Reverse Transcription
• Starting Material: Cells

• Format: Kit
• Reaction Speed: 50 min.
• Optimal Reaction Temperature: 50°C
• Ribonuclease H Activity: Reduced
• Final Product Type: First-Strand cDNA
• Reaction Format: Separate components
• Size (Final Product): Up to 4.5 kb

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