T7 RNA Polymerase, GMP-Grade (GMP-T7P-EE101), DMF #037660
Catalog: GMP-T7P-EE101
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| Product Description | T7 RNA Polymerase (T7RNAP) is the key enzyme for in vitro transcription (IVT) of mRNA. T7 Polymerase is a DNA-dependent polymerase that efficiently incorporates standard or modified nucleoside triphosphates (NTPs) into RNA transcripts.
T7RNAP initiates transcription at the G* in the classic T7 promoter sequence-taatacgactcactataG*GG. T7 Polymerase is highly specific to its promoter sequence. When using the GAG cap1-Analog for co-transcription, taatacgactcactataA*GG is recommended. Note: GMP T7 RNA Polymerase requires the cofactor Mg2+ for mRNA synthesis. We recommend restriction enzymes that produce blunt ends or 5'-overhangs. We've filed our GMP T7RNAP with the FDA Drug Master Files (DMF) and its been assigned DMF #037660. |
| Synonyms | T7RNAP; T7 Polymerase; T7 RNAP; GMP T7 Polymerase; GMP T7 RNA Polymerase; GMP T7RNAP |
| Application | mRNA Synthesis In vitro transcription (IVT) RNA analysis Radiolabeled RNA probe preparation Hybridization RNase protection assay |
| Concentration | 50U/µL |
| Unit Definition | One unit is defined as the amount of enzyme required to incorporate 1 nmol ATP into acid-insoluble material in a total reaction volume of 50µL in 1 hour at 37°C. |
| Source | Expressed in an E.coli strain that carries the T7 RNA Polymerase gene. |
| Molecular Weight | 98 kD |
| Quality Standards | Activity (Molecular Beacon): ≥ 50 kU/mL
Purity (SEC-HPLC): ≥ 95% Residual DNase: Negative Residual RNase: Negative Residual Protease: Negative Endotoxin: ≤ 10 EU/mL Residual Host Cell DNA: ≤ 100 pg/mg Residual Host Protein: ≤ 20 ng/mg Residual Heavy Metal: ≤ 10ppm Residual Nickel Salt: ≤ 10 ppm Bioburden: ≤ 1 CFU/10mL |
| Form | Liquid |
| Shipping | Shipped with dry ice |
| Stability And Storage | -20±5°C for 24 months. Avoid repeated freeze-thaw cycles. |
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Figure 1. Three plasmids (lane 1 with poly A tail, lane 2 and 3 without poly A tail) were used as templates, and the reaction was performed for 2 hours at 37°C in a 20µL system. The transcription length was 2000 nt, 4000 nt, and 2000 nt, respectively. The above mRNA was not purified. |
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Figure 2. Detection of T7 RNA Polymerase by molecular beacon. When the molecular beacon and transcript specifically bind, the conformation changes and the fluorescence intensity changes. Three batches of T7 RNA Polymerase were detected, and the slope was similar across all batches, indicating stable activity across batches. |
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Figure 3. Yield was assessed via nanodrop and Agilent 4150, and purity was assessed by CE. Overall, yield and purity of KACTUS T7 was comparable to leading competitors. |
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