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Cat. No.: CLRKU-25 (for 25T)

Cat. No.: CLRKU-100 (for 100T)

Cat. No.: CLRKU-500 (for 500T)

Description

Colorimetric LAMP/RT-LAMP Assay Kit with UDG is designed for the amplification of DNA through direct or reverse transcription loop-mediated isothermal amplification (LAMP), and the detection of target DNA or RNA in samples by observing color changes in the reaction system. The Bst enzyme in this kit possesses excellent reverse transcriptase activity, making it suitable for detecting both DNA and RNA samples. The kit utilizes dU incorporation and UDG enzyme to prevent contamination of isothermal amplification products. It offers quick, efficient, and highly sensitive detection of target DNA or RNA, employing a visual color change technology that allows results to be determined by the naked eye without the need for electrophoresis. With specially designed isothermal amplification primers, this kit can detect the presence of pathogens and microorganisms in samples, such as specific pathogens or microbial contamination in biological samples.

Isothermal amplification technology is an in vitro nucleic acid amplification technique that maintains a constant temperature throughout the reaction process. Rapid nucleic acid amplification is achieved by adding enzymes with different activities and their respective specific primers. This kit uses loop-mediated isothermal amplification (LAMP) technology, characterized by designing 4-6 specific primers targeting different regions of the target gene, using strand-displacing DNA polymerase (Bst DNA Polymerase) to initiate DNA synthesis, forming dumbbell-like complementary chains, and further entering a cyclic amplification stage through continuous strand displacement. The final amplified product is a mixture of various stem-loop structures and cauliflower-like structures of DNA. The LAMP reaction can be completed in 30-60 minutes under isothermal conditions (e.g., 60-65ºC), offering a simple, rapid, highly sensitive, and specific alternative to conventional PCR, which does not require thermal denaturation of the template, temperature cycling, electrophoresis, or UV observation.

Components

• LAMP Master Mix with UDG (2X)
• Bst DNA Polymerase
• Positive Control
• Nuclease-free Water
• Mineral Oil (Optional)

Features

• This kit offers high sensitivity and a short reaction time. Its sensitivity is 2-5 orders of magnitude higher than traditional PCR methods, with a detection limit of approximately 20-200 copies/μl based on positive plasmid calculations. The detection reaction can be completed in just 1 hour.
• The kit employs visual color change technology, eliminating the need for electrophoresis. Results can be judged by observing the color change with the naked eye. After rapid and extensive amplification of the target DNA fragments in the sample, the optimized indicator dye in the reaction tube changes from violet or bluish-purple to sky blue or deep sky blue, indicating the presence of the target DNA.
• This kit utilizes anti-contamination technology to effectively prevent contamination. By incorporating dU and thermolabile UDG enzymes, the amplified products include dU, and a pre-incubation at 30ºC for 5-15 minutes before isothermal amplification can effectively eliminate product contamination. During isothermal amplification, the thermolabile UDG enzyme is inactivated, thus not interfering with subsequent isothermal amplification detection.
• The kit contains Bst enzyme, UDG enzyme, PCR Buffer, dNTPs, dUTP, and visual dyes. Simply add the appropriate specific primers, the sample to be tested, and water to detect the target DNA/RNA. The Bst enzyme in this kit has good reverse transcriptase activity and can efficiently synthesize cDNA at 60-65ºC, thus eliminating the need for a separate reverse transcription step for RNA samples.
• The kit includes a Positive Control to ensure the kit functions correctly. The Positive Control contains a positive DNA template and corresponding primers. The target DNA for detection is GFP, and the Positive Control includes the corresponding primers and GFP DNA. It is recommended to set up a Positive Control in each test.

Storage 

Store at -20ºC for one year. The LAMP Master Mix with UDG (2X) should be stored protected from light. Avoid repeated freeze-thaw cycles as much as possible.

Precautions

• Ensure the reagents are completely thawed before use. Mix gently by inverting the tube to avoid creating bubbles.
• Testing has shown that the product can withstand up to 10 freeze-thaw cycles without significantly affecting performance. However, repeated freeze-thawing should still be minimized as it may degrade product performance.
• Isothermal amplification is an extremely sensitive detection method. Perform tests in a standard PCR laboratory. Avoid contamination of the isothermal amplification reaction setup area with possible amplification products. Although this kit employs UDG enzyme anti-contamination technology, do not open PCR tube caps. Seal the isothermal amplification products and treat them according to post-amplification product requirements to prevent contamination of the laboratory environment due to aerosolized high-concentration amplification products.
• This product contains a color indicator. If the color of the sample after addition differs significantly from the negative or positive control, dilute the sample or use a DNA extraction kit to process the sample, or adjust the sample's pH to near neutral.
• This isothermal amplification system requires high concentrations of divalent ions; samples should not contain high concentrations of EDTA or other metal ion chelators.
• It is recommended to use filter pipette tips to prepare the isothermal amplification system to minimize contamination leading to false positives.
• This product is for scientific research use by professionals only. It is not intended for clinical diagnosis or treatment, food, or pharmaceutical use, and should not be stored in ordinary residential areas.
• For your safety and health, please wear lab coats and disposable gloves when handling.

Related: Fluorescent LAMP/RT-LAMP Assay Kit with UDG

Only for research and not intended for treatment of humans or animals

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