GeneChip® Two-Cycle cDNA Synthesis Kit (Manufactured by Invitrogen for Affymetrix)
The GeneChip® Two-Cycle cDNA Synthesis Kit is uniquely configured and tested for
GeneChip target labeling. It provides all necessary reagents in one convenient
kit for performing two cycles of double-stranded cDNA synthesis containing the
T7 promoter sequence from 10 to 100 ng of total RNA.
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.
Kit Components
Component name
Concentration
Volume
Quantity
T7-Oligo(dT) Primer, 50 µM
50 µM
120 µL
1
5X 1st
Strand Reaction Mix
5X
180 µL
1
DTT, 0.1M
0.1M
90 µL
1
dNTP, 10 mM
10 mM
200 µL
1
SuperScript™ II
200 U/µL
60 µL
1
RNase Inhibitor
40 U/µL
45µL
1
5X 2nd
Strand Reaction Mix
5X
900 µL
1
Random Primers
3 µg/ µL
60 µL
1
MgCl2,
1M
1M
60 µL
1
E.coli DNA Polymerase
I
10 U/µL
200 µL
1
RNase H
2 U/µL
55µL
1
T4 DNA Polymerase
5 U/µL
60µL
1
RNase-free Water
9.5 mL
1
The Two-Cycle cDNA Synthesis Kit is also available as part of the complete Two-Cycle Target Labeling and Control Reagents package.
