SuperScript® Indirect cDNA Labeling System

Catalog number: L101402

Invitrogen™  Related applications: Microarray Analysis | Nucleic Acid Labeling & Oligo Synthesis

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Description

The SuperScript® Indirect cDNA Labeling System is an array labeling kit based on proven methods of indirect cDNA labeling. The optimized system provides:

• SuperScript® III RT for generating high cDNA yields
• A proprietary nucleotide mixture to increase signal intensity
• A convenient kit format that saves valuable time

In this indirect labeling method, mRNA or total RNA is reverse transcribed using SuperScript® III RT, incorporating amino-modified dUTP and dATP into the synthesized cDNA. The template RNA is then degraded by base hydrolysis, and the reaction is neutralized with acid. The amino-modified cDNA is then purified to remove unincorporated nucleotides, primers, and buffers. In the second step, the modified cDNA is coupled with the active form of a fluorescent dye. The fluorescently labeled cDNA is purified with a S.N.A.P.™ spin column to remove any unreacted dye. The resulting fluorescently labeled cDNA is ready for hybridization to microarrays (Figure 1).

The SuperScript® Indirect cDNA Labeling System provides a complete system that includes both the core kit reagents (except dyes) and a purification module. The SuperScript® Indirect cDNA Labeling Module includes just the core kit reagents. Both provide the flexibility to use fluorescent nucleotides of your choice.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Label or Dye: Alexa Fluor™ 647, Cy™3, Cy™5, Alexa Fluor™ 555
Product Size: 30 reactions
Final Product: cDNA (Labeled)
Labeling Method: Indirect Labeling
Labeling Target: cDNA
Detection Method: Fluorescent
Includes Label or Dye: No
Sample Type (General): RNA

Contents & storage

The SuperScript® Indirect cDNA Labeling Module includes SuperScript® III Reverse Transcriptase, oligo(dT)20- VN (2.5 µg/µl), random primers, control RNA ladder (0.5 g/µl), 5X first-strand buffer, 0.1 M DTT, 10 mM dNTP mixture, RNaseOUT™ Recombinant Ribonuclease Inhibitor (40 U/µl), DEPC-treated water, DMSO (Labeling Grade), 2X coupling buffer, 3M NaAc, pH 5.2, and glycogen (20 mg/ml). The SuperScript® Indirect cDNA Labeling System includes all of the components listed for the Labeling Module, plus S.N.A.P.™ column, collection tubes, loading buffer, wash buffer, amber collection tubes. Store all Labeling Module components at -20°C. Store S.N.A.P.™ columns, collection tubes, loading buffer, wash buffer, and amber collection tubes at room temperature. Guaranteed stable for 6 months when properly stored.

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