Medix Biochemica Isotherm3G DNA Polymerase User Manual
- July 4, 2024
- Medix Biochemica
Table of Contents
Medix Biochemica Isotherm3G DNA Polymerase
Description
Isotherm3G DNA Polymerase is a mesophilic DNA polymerase, which synthesizes DNA from both DNA and RNA templates with a high strand displacement activity. It exhibits 5′ to 3′ polymerase activity but lacks any nuclease activity. Isotherm3G DNA Polymerase is the next generation of the original Isotherm DNA polymerase with further improved reverse transcription activity.
The product components have been optimized and are perfectly suited for loop- mediated isothermal amplification (LAMP) at constant temperature. The optimal reaction temperature is 65°C. However, the enzyme is also active at lower and higher temperatures (60–68°C). The enzyme can be inactivated at temperatures higher than 70°C. Addition of an intercalating dye allows the reaction to be monitored using a real-time PCR instrument. Isotherm3G DNA Polymerase is not suitable for use in PCR.
Isotherm3G DNA Polymerase utilizes an aptamer based warm-start feature to prevent false amplification at low temperatures. For more information, please contact us.
Kit Components
| Component | **S pack*** | **M pack*** |
|---|---|---|
| Isotherm3G DNA Polymerase (8 U/µL) | 1 x 200 µL | 1 x 1 mL |
| 10x Isotherm3G reaction buffer | 1 x 1 mL | 3 x 1 mL |
*Other pack sizes, bulk orders and customization are available upon request.
Storage and Shipment
Transport with ice pack. The reagents should be stored at -20°C upon arrival. The reagents are stable until the expiration date if stored correctly.
Reaction Master Mix Set-Up for LAMP Assay
The recommended reaction master mix set-up for a 25 µL LAMP reaction volume is shown in the table below. Reactions should be setup on ice. Mix and spin down all solutions carefully before use. After the preparation of the master mix, incubate at 65°C for 30– 60 minutes.
To get optimal performance, a temperature gradient experiment (suggested is 60–68°C) and titration of Mg 2+ concentration (suggested is 4–10 mM final) can be performed.
| Reagent | Volume (µL) | Final concentration |
|---|---|---|
| Isotherm3G DNA Polymerase (8 U/µL) | 1 | 0.32 U/µL |
| 10x Isotherm3G reaction buffer | 2.5 | 1x |
| *Primer mix | Variable | Variable |
| dNTPs (10 mM) | 3.5 | 1.4 mM |
| Magnesium chloride (100 mM) | 1.625 | 6.5 mM |
| DNA/cDNA/RNA template | Variable | Variable |
| Nuclease-free Water | Up to 25 µL final volume | |
| Total volume | 25 µL |
*Primer mixes and concentrations depend strongly on the employed isothermal reaction method. Typical primer concentration for LAMP: 1.6 µM FIP/BIP primers, 0.2 µM F3/B3 primers, 0.4 µM LoopF/B primers.
Technical information and support
For technical enquiries or assay development support, please contact us via e-mail at: mdx@medixbiochemica.com.
Additional information and technical resources are available on our website at: info.medixbiochemica.com/resources.
CUSTOMER SUPPORT
myPOLS Biotec GmbH
Byk-Gulden-Strasse 2
78467 Konstanz, Germany
VAT no. DE294545185
www.medixbiochemica.com
www.mypols.de
mdx@medixbiochemica.com
Legal disclaimer
myPOLS Biotec part of Medix Biochemical group products
meet their specifications if transported, stored, and used
according to the instructions. Medix Biochemica’s products
may not be used or reproduced without Medix
Biochemica’s written permission.
References
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