Description
This product is a bacteriophage T7 RNA polymerase derived from recombinant expression in E. coli. It is a T7 RNA polymerase obtained by mutation after modification and optimization of wild-type T7, which can significantly reduce the dsRNA content. Double-stranded DNA containing the T7 promoter sequence is used as a template and NTP is used as a substrate to synthesize RNA complementary to the reverse single-stranded DNA downstream of the promoter. Double-stranded linear blunt-end or 5' protruding end DNA can be used as a substrate template for T7 RNA polymerase, so linear plasmids and PCR products can be used as templates for in vitro RNA synthesis.
Specifications
|
Source |
Recombinant E. coli |
|
Optimum temperature |
37℃ |
|
Activity |
1000 U/μL |
|
Residual RNase |
Negative |
|
Storage buffer |
50 mM Tris-HCl, 1 mM EDTA, 10 mM DTT, 100 mM NaCl, 0.1% Triton X-100, 50% (v/v) glycerol, pH7.9@25℃ |
|
Activity unit definition |
The amount of enzyme required to incorporate 1 nmol of [3H] GMP into the acid-insoluble precipitate within 1 hour at 37℃ and pH8.0 is defined as one activity unit. |
Components
|
Components No. |
Name |
10636ES50 (50 KU) |
10636ES60 (500 KU) |
10636ES72 (1 MU) |
10636ES86 (10 MU) |
|
10636 |
HieffTM T7 RNA Polymerase (low dsRNA, 1000 U/μL) |
50 μL |
500 μL |
1 mL |
10 mL |
Storage
This product should be stored at -25~-15℃ for 1 year.
Figures
1. Customer feedback 1
Experimental Design:
|
Parameter |
Condition |
|
Objective |
To compare application performance between WT T7 and Low-dsRNA T7 |
|
Reaction Volume |
20 µL |
|
Template Length |
2 kb |
|
Test Samples |
WT T7 & Low-dsRNA T7 RNA polymerase |
|
Reaction Temperature |
27 °C |
|
Reaction Time |
2 h |
Results:
Table 1. Comparison of dsRNA Content and Yield Between WT T7 and Low-dsRNA T7
|
Enzyme Type |
dsRNA Content (%) |
Integrity (%) |
Reaction Scale |
|
WT T7 |
0.0030% |
92.5 |
180 µg (20 µL) |
|
Low-dsRNA T7 |
0.0003% |
93.0 |
180 µg (20 µL) |
Result Summary:
Low-dsRNA T7 reduced dsRNA content by approximately 10-fold while maintaining comparable RNA yield and integrity relative to WT T7.
2. Customer feedback 2
Table 2. Performance Comparison Under Different GAG Final Concentrations Yield (mg/mL)
|
GAG Final Concentration (mM) |
Yield (mg/mL) |
Integrity (%) |
Capping Efficiency (%) |
Transfection Efficiency (%) |
||||||||
|
|
WT |
Yeasen |
supplier A* |
WT |
Yeasen |
supplier A* |
WT |
Yeasen |
supplier A* |
WT |
Yeasen |
supplier A* |
|
10 |
12.14 |
11.376 |
12.103 |
88.1 |
92.5 |
92.3 |
96.7 |
100 |
100 |
44.60 |
33.63 |
30.76 |
|
5 |
11.82 |
10.716 |
11.337 |
82.2 |
91.1 |
91.5 |
98.3 |
100 |
100 |
42.30 |
32.78 |
32.37 |
|
2.5 |
11.31 |
10.716 |
11.066 |
82.9 |
91.7 |
91.8 |
98.8 |
97.3 |
97.8 |
39.78 |
31.68 |
32.59 |
|
1.5 |
11.98 |
— |
— |
78.9 |
— |
— |
97.7 |
— |
— |
— |
— |
— |
|
1 |
11.75 |
— |
— |
80.0 |
— |
— |
90.5 |
— |
— |
— |
— |
— |
|
0.5 |
11.65 |
— |
— |
80.9 |
— |
— |
70.1 |
— |
— |
— |
— |
— |
Result Summary:
Effects of cap analog concentration on IVT performance. Reactions were carried out in a 20 µL system at 27 °C for 2 h using different T7 enzymes. RNA yield remained stable as cap analog input decreased, while integrity and capping efficiency showed a gradual decline. A cap analog concentration of ≥2.5 mM is recommended for optimal performance.
Documents:
Safety Data Sheet
Manuals
Paiement et sécurité
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Enquête
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FAQ
Le produit est destiné à des fins de recherche uniquement et n'est pas destiné à un usage thérapeutique ou diagnostique chez l'homme ou l'animal. Les produits et le contenu sont protégés par des brevets, des marques déposées et des droits d'auteur appartenant à Yeasen Biotechnology. Les symboles de marque indiquent le pays d'origine, pas nécessairement l'enregistrement dans toutes les régions.
Certaines applications peuvent nécessiter des droits de propriété intellectuelle tiers supplémentaires.
Yeasen se consacre à la science éthique, estimant que nos recherches doivent répondre à des questions cruciales tout en garantissant la sécurité et les normes éthiques.