规格
|
Hot Start:
|
No |
Polymerase:
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T7 RNA Polymerase |
Promoter:
|
T7 |
Shipping Condition:
|
Wet Ice |
Quantity:
|
2500 units |
储存
|
T7 RNA Polymerase is supplied with a vial of 5X reaction buffer [200 mM Tris-HCl (pH 8.0), 40 mM MgCl2 , 10 mM spermidine-(HCl)3 , 125 mM NaCl], vial of 100 mM DTT. Store at -20°C. |
T7 RNA Polymerase is a DNA-dependent RNA polymerase that has a high specificity for bacteriophage T7 promoter sequences. The enzyme synthesizes large quantities of RNA from DNA inserted into a transcription vector downstream from a T7 promoter. A T7 RNA Polymerase technical bulletin is available.
Application: Synthesis of labeled and unlabeled RNA transcripts (1).
Source: Purified from E. coli expressing the T7 RNA Polymerase gene on a plasmid.
Performance and Quality Testing: 3´ and 5´ exodeoxyribonuclease, ribonuclease, and DNA nicking assays; performance in a
transcription reaction.
Unit Definition: One unit hydrolyzes 1 nmol of ribonucleotide into acid-precipitable material in 1 h at 37°C using a T7 transcription
vector as template.
Unit Reaction Conditions: 40 mM Tris-HCl (pH 8.0), 25 mM NaCl, 8 mM MgCl2 , 2 mM spermidine-(HCl)3 , 5 mM DTT, 0.4 mM ATP, 0.4 mM CTP, 0.4 mM GTP, 0.4 mM UTP, 1 µCi [3H]GTP, 1 µg Sph I-cut pT7L13, and enzyme in 50 µl for 10 min. at 37°C.