Nucleic Acid Test
Reverse Transcription Kit
● Product Name: Magigen Thermostable Reverse Transcriptase III - RT enzyme For RT-PCR Test
● Catalog# : A003M
● Size: 500µl , 200U/ µl
● Reverse Transcriptase Price: 903 USD
● Contents
Composition Vol
5× Reaction Buffer 1 1 ml×3
thermostable reverse transcriptase III 500 µl×1
100 mM DTT 500 µl×2
ddH2O 1.5 ml×2
● Magigen RT enzyme Features
Magigen Thermostable Reverse Transcriptase, RTase III, is a significant advancement over popular Moloney Murine Leukemia Virus reverse transcriptase (M-MLV reverse transcriptases), and expressed using gene recombination technology. RNase H activity has been removed completely, and different species of reverse transcriptase fragments are chimerized in different structural regions. The product has excellent thermal stability, and the residual activity can reach almost 100% at 65 ℃.
● Unit Definition
One unit is defined as the amount of enzyme that will incorporate 1 nmol of dTTP into acid-insoluble material in 10 minutes at 37°C, using poly(rA) as a template, oligo(dT) as a primer.
● Storage Conditions and Expiration
Sealed at-20℃, valid for 12 months.
● How to use thermostable Reverse Transcriptase III
Example: first strand cDNA synthesis
Reaction System Vol ( µl)
Thermostable RTase III 1
5× Reaction Buffer 1 4
100 mM DTT* 2
RNase Inhibitor（20-40 U/µl） 1
dNTP Mix ( 25 mM each) 1
primer 10μM-250μM
template 1pg-100ng
ddH2O Up to 20-25
* A final concentration of 10 mm is recommended
● Reaction steps
1. gently mix the reaction system;
2. recommended reaction procedure:
Temperature Time
25℃1 2min
50℃2 30min
85℃ 2min
*1. This step is only required when using random hexmers;
* 2. For templates with complex structure or high GC, the reaction temperature can be increased to 55 ℃;
For cDNA fragments longer than 3 KB, the reaction temperature can be increased to 65 ℃, which helps to improve the yield.
● RTase III Kit Application
The obtained cDNA can be directly used for 2nd strand cDNA synthesis or PCR amplification.
● Heat Inactivation
80°C for 20 minutes
● Note
1. The optimal temperature range of magicscript thermostable reverse transcriptase III is 37-65 ℃. With the increase of temperature, the residual activity will remain more than 95%; Its RNaseH activity is completely lost, and the length of its inversion product can exceed 12 KB. At the same time, the tolerance to inhibitors such as high salt, fat and heparin was significantly enhanced.
2. The thermostable reverse transcriptase III is not recommended in the one-step reaction because it contains a special active domain. Although it improves the reverse transcription efficiency, thermostability and accuracy, it will affect the polymerase activity in the presence of metal ions, thermostable reverse transcriptase III and DNA dependent polymerase at the same time.
Researchers can increase the reaction temperature to obtain better experimental results when reverse transcribing complex templates, and can optimize various parameters according to the experimental design or use the reaction system explored by researchers.
● Notice
Magigen Reverse Transcription Kit is a high quality product with a good price.
The Reverse Transcription Kit price is subject to change without prior notice due to currency fluctuation.
The Reverse Transcription Kit price is FOB price, not include tariffs, freight, insurance and other expenses. Some products need to be delivered by dry ice. Please consult the sales manager for details before purchasing.
All Pictures shown are for illustration purpose only and shall not form part of any offer, contract.
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