Описание
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
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Расследование
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