What components does the GeneChip® One-Cycle cDNA Synthesis Kit contain?
The One-Cycle cDNA Synthesis Kit contains all necessary reagents (see the following table) to carry out the cDNA synthesis reaction for thirty samples in a "standard" assay for GeneChip arrays. From total RNA starting materials (1 μg to 15 μg), using this kit, sufficient double-stranded cDNA can be synthesized, purified on the cDNA Cleanup Column, and used as template in the subsequent in vitro transcription labeling reaction.
Component Name |
Concentration |
Volume |
Quantity |
|---|---|---|---|
| T7-Oligo(dT) Primer, 50 µM | 50 μM | 120 μL | 1 |
| 5X 1st Strand Reaction Mix | 5X | 120 μL | 1 |
| DTT, 0.1M | 0.1M | 60 μL | 1 |
| dNTP, 10 mM | 10 mM | 120 μL | 1 |
| SμperScript™ II | 200 U/μL | 60 μL | 1 |
| 5X 2nd Strand Reaction Mix | 5X | 900 μL | 1 |
| E.coli DNA Ligase | 10 U/μL | 30 μL | 1 |
| E.coli DNA Polymerase I | 10 U/μL | 120 μL | 1 |
| RNase H | 2 U/μL | 30 μL | 1 |
| T4 DNA Polymerase | 5 U/μL | 60 μL | 1 |
| EDTA, 0.5M | 0.5M | 300 μL | 1 |
| RNase-free Water | 3.1 mL | 1 |