SuperScript® III CellsDirect™ cDNA Synthesis Kit

产品概述

描述

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.

Advantages of the SuperScript® III CellsDirect cDNA 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 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 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.

Note: For real-time quantitative RT-PCR (qRT-PCR) from cell lysate, we recommend the Cells-to-CT™ family of products for RNA extraction. These kits include reagents and protocols that have been specifically optimized for real-time qRT-PCR.

规格

Reverse Transcriptase:	SuperScript® III
Downstream Application:	Reverse Transcriptase PCR (RT-PCR), Real-Time Quantitative PCR (qPCR)
Reaction Format:	Separate Components
Sample Type (General):	Cells
Product Size:	25 reactions
Optimal Reaction Temperature:	50° C

内容及储存

The SuperScript® III CellsDirect cDNA Synthesis Kit includes SuperScript® III RT, RNaseOUT™ Ribonuclease Inhibitor, Oligo(dT)20 primers, DNase I, DNase I buffer, EDTA, dNTP mix, 5X first-strand buffer, 0.1 M DTT, RNase H, control RNA and primers, and resuspension buffer. Store all components at -20°C. Guaranteed stable for 6 months when properly stored.

文档

手册和实验方案     

• SuperScript III CellsDirect cDNA Synthesis System

cDNA Synthesis Directly from Cells Using SuperScript™ III CellsDirect cDNA Synthesis System |

化学品安全技术说明书(MSDS)

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• 引用及参考文献

  Transfer of a human chromosomal vector from a hamster cell line to a mouse embryonic stem cell line.
  Paulis M, Bensi M, Orioli D, Mondello C, Mazzini G, D'Incalci M, Falcioni C, Radaelli E, Erba E, Raimondi E, De Carli L, 
  Stem Cells (2007) 25:2543-2550
  Product usage: SuperScript III CellsDirect cDNA Synthesis Kit was used to produce total cDNA from embryonic stem cells. The retention of the pluripotent phenotype of hybrid cells was verified by RT-PCR for the expression of the early embryonic developmental markers Oct4 and Nanog.
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产品文

• CellsDirect - endpoint