This reagent kit is based on LAMP, a new nucleic acid amplification technology. Real time monitoring of LAMP amplification is carried out by adding SYBR Green dye. It requires a fluorescence quantitative PCR instrument or a constant temperature amplification fluorescence detector, and the requirements for primers are relatively high. High specificity primers can better avoid the occurrence of false positives.
LAMP is a novel nucleic acid amplification technique that uses 4 or 6 primers that can recognize 6 specific regions on the target gene. It relies on the strong strand displacement activity of Bst DNA polymerase and can amplify DNA 109-1010 times within 30-60 minutes.
Bst DNA polymerase is a part of Bacillus stearothermophilus DNA polymerase, which has strong strand displacement activity and 5 '→ 3' DNA polymerase activity. Therefore, it can be applied to LAMP (Loop mediated isothermal amplification) detection experiments. This product contains Bst DNA polymerase 4.0, which can amplify both DNA and RNA molecules directly.
Kit features:
·The hot start enzyme adopts a proprietary modification method, with a 30 second hot start;
·Dry powder products, performance and stability;
·Excellent amplification performance, strong tolerance to impurities in the sample;
·High sensitivity, capable of detecting low abundance genes;
·Suitable for all real-time quantitative PCR instruments.
Usage:
1. Dissolved freeze-dried products
Lyophilized LAMP SYBR Green Reagent is a freeze-dried powder containing Bst DNA polymerase 4.0, dNTPs, etc. Before dissolution, the product can be stored for a long time at -20 ℃ and transported at room temperature without any degradation in performance. After dissolving each bottle with 0.75mL LAMP Buffer before use, it can be used immediately. The remaining reagents can be stored at -20 ℃ for 1 month and stored for a long time below -60 ℃.
2. Configure reaction system
Prepare 20 according to the following components μ PCR reaction system
2.The setting of the fluorescence quantitative PCR instrument LAMP SYBR Green can be detected using a standard quantitative PCR instrument (or a constant temperature fluorescence device).
Using a quantitative PCR instrument for LAMP SYBR Green amplification program:
Step 1: 60 ℃ for 10s
Step 2: Collect the signal at 60 ℃ for 50 seconds and cycle 25 times
The total reaction time is 25 minutes.
Note: It is prohibited to open the reaction tube cover after the LAMP reaction is completed, otherwise it may cause aerosol pollution. Once aerosol pollution occurs, please use our DNA aerosol pollution remover (product number: MT3001) for environmental cleaning.
