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.