What are the major differences and improvements comparing the new Two-Cycle cDNA Synthesis Kit with the Affymetrix® Small Sample Target Labeling Version II Protocol?
The Two-Cycle cDNA Synthesis Kit was developed primarily based on SSTLvII, with some additional optimization focusing on a more streamlined procedure. More information on these changes is summarized as follows:
- The new Two-Cycle cDNA Synthesis Kit protocol utilizes larger reaction volume for the 1st-cycle cDNA synthesis step, allowing more flexibility in the starting material concentration and reducing potential pipetting variations.
- The new Two-Cycle cDNA Synthesis Kit protocol eliminates the cDNA cleanup step in the 1st cycle, increasing ease of use.
- The GeneChip® Sample Cleanup Module is used for all cDNA and cRNA cleanup steps, replacing all previously recommended precipitation or speed vacuum protocols.
- Overnight incubations are recommended for both IVT steps following the 1st- and 2nd-cycle cDNA synthesis, reducing the total target labeling time to 2½ days.