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Invitrogen™ SuperScript™ Indirect cDNA Labeling Module, no purification columns
Description
Includes
The SuperScript™ Indirect cDNA Labeling Module includes SuperScript™ III Reverse Transcriptase, oligo(dT)20- VN (2.5μg/μL), random primers, control RNA ladder (0.5g/μL), 5X first-strand buffer, 0.1M DTT, 10mM dNTP mixture, RNaseOUT™ Recombinant Ribonuclease Inhibitor (40U/μL), DEPC-treated water, DMSO (Labeling Grade), 2X coupling buffer, 3M NaAc, pH 5.2, and glycogen (20mg/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.
The SuperScript Indirect cDNA Labeling System provides a complete system that includes both the core kit reagents (except dyes) and a purification module. Modules do not include all components. The SuperScript Indirect cDNA Labeling Module includes just the core kit reagents. Both provide the flexibility to use fluorescent nucleotides of your choice. The full SuperScript Indirect cDNA Labeling System is an array labeling kit based on proven methods of indirect cDNA labeling. Provides the flexibility to use fluorescent nucleotides.
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.
- SuperScript III RT for generating high cDNA yields
- A proprietary nucleotide mixture to increase signal intensity
- A convenient kit format that saves valuable time
DNA and RNA Purification and Analysis, DNA Labeling, Expression Array Labeling, Gene Expression Analysis and Genotyping, Microarray Analysis, Nucleic Acid Labeling and Oligo Synthesis
Specifications
Specifications
| Product Type | cDNA Labeling Module |
| Detection Method | Fluorescence |
| Content And 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. |
| Final Product Type | cDNA (Labeled) |
| Label or Dye | Alexa Fluor™ 555, Alexa Fluor™ 647, Cy™3, Cy™5 |
| Includes Label or Dye | No |
| Labeling Target | cDNA |
| Labeling Method | Indirect Labeling |
| Sample Type | RNA |
| Product Line | SuperScript |
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Frequently Asked Questions (FAQs)
Yes, the amino-labeled cDNA should be stable if stored precipitated in ethanol at -20 degrees C.
The ratio of nucleotides is one of the key features in the optimized performance of this product, and therefore is considered proprietary. Unfortunately, we are unable to provide this information.
We have not used bacterial RNA for the validation of the SuperScript Indirect cDNA Labeling System, however the kit should work reasonably well with few modifications to the protocol.
For total RNA samples, Oligo(dT)20 primers should be used. As the manual suggests, random hexamers should be used only with mRNA or with the RNA ladder, which is included as a control. However, in the case of bacterial RNA, longer random primers should be used instead of the random hexamers included in the kit. 18mers should work well, although the temperature of reverse transcription may need to be slightly adjusted in order to obtain optimal cDNA synthesis. The reason for using longer random primers is that the rRNA in bacterial samples will also be used as template for reverse transcription, and the target cDNA will be more complex than the corresponding cDNA primed with the Oligo(dT)20. Therefore, when doing hybridizations, you can expect some degree of cross-hybridization to occur. Nevertheless, the use of long random 18mers is common practice among microarray users.
For Research Use Only. Not for use in diagnostic procedures.