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Cat. No.: FLRKU-25 (for 25T)

Cat. No.: FLRKU-100 (for 100T)

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

Description

Fluorescent LAMP/RT-LAMP Assay Kit with UDG is designed to amplify DNA through direct or reverse transcription loop-mediated isothermal amplification (LAMP), and to determine the presence of target DNA or RNA in samples by analyzing the fluorescence curve on a quantitative PCR instrument. The Bst enzyme in this kit exhibits excellent reverse transcriptase activity, enabling it to detect both DNA and RNA samples. This kit incorporates dU and UDG enzymes, effectively preventing false positives caused by contamination of isothermal amplification products. It offers quick, efficient, and highly sensitive detection of target DNA or RNA. With specially designed isothermal amplification primers, this kit can detect the presence of pathogens or microorganisms in samples, such as identifying infections by 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. Rapid nucleic acid amplification is achieved by adding enzymes with different activities and their respective specific primers. This kit utilizes loop-mediated isothermal amplification (LAMP) technology, characterized by designing 4-6 specific primers targeting six regions of the target gene. Strand-displacing DNA polymerase (Bst DNA Polymerase) initiates DNA synthesis, forming dumbbell-like complementary chains. These enter a cyclic amplification stage through continuous strand displacement, producing a mixture of DNA with various stem-loop structures and cauliflower-like structures as the final amplified products. LAMP reactions can be completed within 30-60 minutes under isothermal conditions (e.g., 60-65ºC). Compared to conventional PCR, LAMP does not require thermal denaturation of the template, temperature cycling, electrophoresis, or UV observation, offering simplicity, rapidness, high sensitivity, and strong specificity.

Components

• LAMP Fluorescent Mix with UDG (2X)
• Bst DNA Polymerase
• Positive Control
• Nuclease-free Water

Features

• This kit offers high sensitivity and a short reaction time. Its sensitivity is 2-5 times 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 30-60 minutes.
• 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 37ºC 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 SYBR Green dye. 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 target DNA for detection is GFP. The Positive Control includes 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 Fluorescent Mix with UDG (2X) should be stored protected from light and repeated freeze-thaw cycles should be avoided as much as possible.

Precautions

• Ensure the reagents are completely thawed before use. Mix gently by inverting the tube, and avoid creating bubbles during mixing.
• 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 to avoid potential degradation.
• Isothermal amplification is an extremely sensitive detection method. Perform tests in a standard PCR laboratory to 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 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.
• Treat the pipette with DNA removal reagents before and after use to avoid potential 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: Colorimetric LAMP/RT-LAMP Assay Kit with UDG

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

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