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EA0202 LAMP Fluorescent Master Mix

Cat. No. EA0202
Source /
Appearance Liquid
Endotoxin /
Purity /
Consult Immediately
Detail

LAMP Fluorescent Master Mix (Liquid)

Cat#EA0202

Description

LAMP is a novel nucleic acid amplification technique.The use of 4 or 6 primers capable of recognising 6 specific regions on the target gene, relying on the strong strand displacement activity of Bst DNA polymerase, the DNA amplification can reach 109-1010 fold in 30-60 minutes. LAMP Master Mix with Dye is used for the detection of template DNA, providing a rapid one-step solution.

This premix uses an optimised Bst DNA polymerase formulation for rapid ring-mediated isothermal amplification (LAMP) reactions. This premix contains all the necessary components including Enzyme Mix, dNTP, Fluorescent Dye and Reaction Buffer. Simply add primers and templates for isothermal amplification and real-time fluorescence detection. Ready to use amplification results were obtained within 30-50min.

Product Composition

Name

Size (100 T)

2 × LAMP Master Mix with Dye

0.625 mL × 2

Storage Conditions

Transport on dry ice and avoid exposure to frequent temperature changes. It can be stored at -20 °C for a short time and at -80 °C for one year.

Experimental Procedures

1. There will be a slight precipitate after the product is taken out at -20 ℃, it should be thawed and mixed at room temperature until the precipitate disappears, then centrifuged and used.

2. Configure the reaction mixture according to the following components:

Component

Volumes

2 × LAMP Master Mix with Dye

12.5 μL

10 × LAMP Primer Mix*

2.5 μL

Template DNA

X μL

ddH2O

Up to 25 μL

*Recommended final primer concentrations: 1.6 μM FIP/BIP, 0.4 μM LF/LB, 0.2 μMF3/B3. Mix gently and centrifuge.

3. Place the reaction in a fluorescence quantifier at 65 ℃ and collect the fluorescence signal once a minute, set to 35-45cycles.

4. Thermal inactivation condition: 85 ℃ for 5-10 min. (It is generally prohibited to open the cap of the reaction tube after the LAMP reaction, if subsequent electrophoresis and other open-cap operations are carried out, thermal inactivation must be carried out to prevent aerosol contamination).

Notes

1. It is recommended that a negative control be added to each experiment.

2. It is recommended that reagent preparation and template addition be done in different areas to avoid environmental contamination, which may affect the subsequent experiments.

3. It is recommended that the template DNA be added last in order to ensure good replication results.

4. It is recommended that plasmid DNA be added between 1fg-10 ng and genomic DNA between 1pg-100 ng.

5. Recommended primer online design software: recommended http://primerexplorer.jp/e/

Examples of applications

LAMP1.png

LAMP amplification of N gene plasmid