Sự miêu tả
Hifair™ III Reverse Transcriptase is a novel reverse transcriptase engineered from Hieff™ M-MLV (H-) Reverse Transcriptase. Compared to its predecessor, Hifair™ III exhibits significantly enhanced thermostability, tolerating reaction temperatures of up to 65°C, making it ideal for reverse transcribing RNA templates with complex secondary structures. Additionally, this enzyme features increased template affinity, making it highly suitable for reverse transcription of limited samples and low-copy genes. Its ability to synthesize full-length cDNA has also been improved, capable of extending cDNA up to 19.8 kb. This glycerol-free version (600 U/μL) is specifically designed for the preparation of lyophilized formulations and lyophilizable RT-LAMP reagents.
Specifications
|
Cat.No. |
11297ES09 / 11297ES12 / 11297ES75 |
|
Size |
6 KU / 12 KU / 300 KU |
Components
|
Components No. |
Name |
11297ES09 |
11297ES12 |
11297ES75 (300 KU) |
|
11297 |
Hifair™ III Reverse Transcriptase (600 U/µL, Glycerol-free) |
10 μL |
20 μL |
500 μL |
Storage
This product should be stored at 2~8℃ for 6 months.
Applications
Preparation of lyophilized formulations
Lyophilizable RT-LAMP kits
Activity Definition
One unit (U) is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insoluble material using Poly(A)·Oligo(dT) as the template-primer in 10 minutes at 37°C.
Notes
1. This product is for research use only.
2. Please operate with lab coats and disposable gloves，for your safety.
3. Maintain a clean experimental area. Wear clean gloves and a mask during operation. Ensure all consumables are RNase-free to prevent RNase contamination.
4. Perform all operations on ice to prevent RNA degradation.
5. High-quality RNA samples are recommended to ensure efficient reverse transcription.
Instructions
1. Reaction System Setup
|
Cat. No. |
Component |
Volume |
|
13840-A |
10×Bst Buffer |
2.5 µL |
|
13840-C |
100 mmol/L MgSO4 |
2 µL |
|
dNTP Mix (100 mmol/L) |
dNTP Mix (100 mmol/L) |
3.5 µL |
|
dUTP Mix (100 mmol/L)(Optional) |
dUTP Mix (100 mmol/L)（Optional） |
0.35 µL |
|
11297 |
Hifair™ III Reverse Transcriptase (600 U/µL, Glycerol-free) |
0.1 µL |
|
10703 |
Murine RNase inhibitor (200 U/µL, Glycerol-free) |
0.2 µL |
|
13839(Optional) |
Heat Labile UDG (200 U/µL)（Optional） |
0.005 µL |
|
10×Primersa |
10×Primers |
2 µL |
|
13840-Bb |
Bst Ⅱ Plus DNA polymerase (120 U/µL, Glycerol-free) |
0.5 µL |
|
RNase free ddH2O |
RNase free ddH2O |
To 25 µL |
[Note]:
a. 10× Primers: 16 µmol/L FIP/BIP, 2 µmol/L F3/B3, 4 µmol/L Loop F/B each. The primer ratios can be adjusted as needed.
b. Volumes can be adjusted according to specific experimental conditions.
2. Reaction Program Settings:
|
Temperature |
Time |
Function |
|
37℃ |
2 min |
Digestion |
|
65℃ |
30-60 mina |
Reverse Transcription and Isothermal Amplification |
|
85℃ |
10 min |
Enzyme Inactivation |
[Note]:
a. For probe-based methods, fluorescence can be collected directly in the target channel at 1-minute intervals. For dye-based methods, you may additionally add Dye (Cat# 13841), select the SYBR channel, and collect fluorescence at 1-minute intervals.
Documents:
Safety Data Sheet
Manuals
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