GeneChip® One-Cycle cDNA Synthesis Kit (Manufactured by Invitrogen for Affymetrix)
The One-Cycle cDNA Synthesis Kit is uniquely configured and tested
for GeneChip target labeling. It provides all necessary reagents in one convenient
kit for synthesis of double-stranded cDNA containing the T7 promoter sequence
from 1 to 15 µg of total RNA or 0.2 to 2 µg of mRNA.
The resultant cDNA is then purified and serves as a template in the subsequent
in vitro transcription (IVT) reaction for complementary RNA (cRNA) amplification
and biotin labeling to hybridize to GeneChip® arrays.
As part of the completely integrated GeneChip reagent product line for expression
analysis, the GeneChip One-Cycle cDNA Synthesis Kit offers:
•
Optimal results
based on proven, robust protocol
•
Consistent performance
with SuperScript™ II from Invitrogen for reverse transcription
•
Limited license
to Van Gelder/Eberwine patents via Incyte
Kit Components
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
SuperScript™ 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
The One-Cycle cDNA Synthesis Kit is also available as part of the complete One-Cycle Target Labeling and Control Reagent package.
