Beschreibung
T7 High Yield RNA Synthesis Kit (low dsRNA) optimizes the transcription reaction system. The T7 RNA polymerase used in this kit is an engineered and optimized variant of wild-type T7 RNA polymerase, obtained by mutation, which can significantly reduce dsRNA content. Using linear double-stranded DNA containing a T7 promoter sequence as template and NTPs as substrates, it transcribes the DNA sequence downstream of the promoter and efficiently synthesizes single-stranded RNA. Modified nucleotides can be incorporated into the substrates during transcription to prepare biotin- or dye-labeled RNA.
This kit can synthesize both long and short transcripts. With 1 μg of template input, 100–200 μg of RNA can be produced. The synthesized RNA can be used for a variety of downstream applications such as RNA structure and function studies, RNase protection, probe hybridization, RNAi, microinjection, and in vitro translation.
Product Information
|
Catalog No. |
Size |
|
10633ES10 / 10633ES50 / 10633ES60 / 10633ES70 |
10 T / 50 T / 100 T / 500 T |
Component Information
|
Component No. |
Component Name |
10633ES10 |
10633ES50 |
10633ES60 |
10633ES70 |
|
10633-A |
T7 RNA Polymerase Mix (low dsRNA), 20 U/μL |
20 μL |
100 μL |
200 μL |
1 mL |
|
10633-B |
10× Transcription Buffer |
20 μL |
100 μL |
200 μL |
1 mL |
|
10633-C |
ATP (100 mM) |
20 μL |
100 μL |
200 μL |
1 mL |
|
10633-D |
CTP (100 mM) |
20 μL |
100 μL |
200 μL |
1 mL |
|
10633-E |
GTP (100 mM) |
20 μL |
100 μL |
200 μL |
1 mL |
|
10633-F |
UTP (100 mM) |
20 μL |
100 μL |
200 μL |
1 mL |
|
10633-G |
Control DNA Template (500 ng/μL) |
5 μL |
10 μL |
20 μL |
100 μL |
|
10633-H |
DNase I (2 U/μL) |
10 μL |
50 μL |
100 μL |
500 μL |
|
10633-I |
Lithium Chloride Solution |
300 μL |
1.5 mL |
3 mL |
15 mL |
|
10633-J |
RNase-free ddH₂O |
500 μL |
2.5 mL |
5 mL |
25 mL |
Feature
• Extremely high yield—up to 200 μg in 2 hours
• Broad applicability—suitable for transcription of 20 nt to 10,000 nt RNA
• Extremely low dsRNA—significantly reduces dsRNA content during IVT
• Multiple applications—suitable for standard, labeled, and modified RNA synthesis
Applications
1. Under different length applications, both 2K and 4K fragments achieved yields of ~11 mg/mL; with the low dsRNA T7 system, dsRNA content was drastically reduced.
|
Fragment length |
T7 RNA pol |
Yield (mg/mL) |
Integrity (%) |
dsRNA content (ng dsRNA produced from 1 μg RNA) |
|
2K |
T7-WT |
12.4 |
93.4 |
0.01728 |
|
|
T7-low dsRNA |
12.2 |
94.1 |
0.0003 |
|
4K |
T7-WT |
11.4 |
89.3 |
0.4273 |
|
|
T7-low dsRNA |
11.7 |
89.4 |
0.0129 |
2. High integrity
Figure 1: 44–4000 nt RNA analyzed on the Agilent 2100 Bioanalyzer. The electrophoretic bands are single and the peaks are distinct and sharp, indicating high RNA integrity.
Storage
Store at −25°C to −15°C. Shelf life: 1 year.
Documents:
Safety Data Sheet
Manual
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