Manual

Product Number: LAP017

Shipping and Storage

Store at -20ºC, valid for one year. LAMP Master Mix with UDG (2×) must be protected from light. Avoid repeated freeze-thaw cycles as much as possible.

Component

Description

pH-Sensitive Color-Changing Isothermal Amplification Kit (LAMP/RT-LAMP, with UDG) by our company is a kit for amplifying DNA/RNA via Loop-Mediated Isothermal Amplification (LAMP) and detecting the presence of target DNA/RNA in samples through color change of the reaction system. The Bst polymerase in this kit exhibits excellent reverse transcriptase activity, enabling detection of both DNA and RNA templates. This kit incorporates dUTP incorporation and uracil-DNA glycosylase (UDG) technology to prevent contamination from isothermal amplification products, allowing rapid, efficient and highly sensitive detection of target DNA/RNA in samples. It adopts visual colorimetric technology, eliminating the need for electrophoresis, and results can be judged by naked-eye observation of color change. By designing isothermal amplification primers independently, this kit can be used to detect the presence of pathogens, microorganisms, etc. in samples, such as specific pathogen infections or microbial contaminations in biological samples.

Isothermal amplification technology is an in vitro nucleic acid amplification technology that maintains a constant temperature throughout the reaction process and achieves rapid nucleic acid amplification by adding enzymes with different activities and their respective specific primers. This kit employs Loop-Mediated Isothermal Amplification (LAMP), which is characterized by designing 4-6 specific primers targeting different regions of the target gene, using strand-displacement DNA polymerase (Bst DNA Polymerase) to initiate DNA synthesis, forming dumbbell-shaped complementary strands, and further entering the cyclic amplification stage through continuous strand displacement. The final products of amplification are a mixture of DNA with multiple stem-loop structures and cauliflower-like structures. LAMP can complete the nucleic acid amplification reaction only by incubating at an isothermal condition (e.g., 60-65℃) for 30-60 minutes. Compared with conventional PCR, LAMP does not require thermal denaturation of templates, temperature cycling, electrophoresis or ultraviolet observation, and has the advantages of simplicity, rapidity, high sensitivity and strong specificity.

Feature

1. High sensitivity and short reaction time: The sensitivity of this kit is about 10 times higher than that of traditional PCR methods, with a detection limit of approximately 20-200 copies/μL calculated using positive plasmids. The detection reaction can be completed in only 30-60 minutes.
2. Visual colorimetric detection: No electrophoresis is required, and results can be judged by naked-eye observation of color change. This kit contains an optimized red indicator dye. After the target DNA/RNA fragments in the sample are rapidly and massively amplified, the liquid in the reaction tube changes from red or rose red to orange-yellow or yellow, indicating a positive result for target DNA/RNA. Compared with pH-Sensitive Color-Changing Isothermal Amplification Kit (LAMP/RT-LAMP, with UDG), which changes from violet or blue-violet to sky blue or deep sky blue, this kit shows more stable color change and more distinct distinction between negative and positive results.

Figure. Detection effect of pH-Sensitive Color-Changing Isothermal Amplification Kit (LAMP/RT-LAMP, with UDG). Left: Negative sample; Right: Positive sample. This figure is for reference only. The colors of negative and positive results may vary slightly between different batches of products. A significant color change between positive and negative samples during actual detection is acceptable.

• Effective anti-contamination technology: This kit uses dUTP and thermolabile UDG enzyme, which allows dUTP to be incorporated into amplification products. Incubation at 37℃ for 5-15 minutes before isothermal amplification can effectively eliminate product contamination during the isothermal amplification process. During isothermal amplification, the thermolabile UDG enzyme will be inactivated, thus not interfering with subsequent isothermal amplification detection.
• All-in-one reaction system: This kit contains Bst polymerase, UDG enzyme, PCR Buffer, dNTPs, dUTP and visual indicator. Only appropriate specific primers, test samples and water need to be added to detect target DNA/RNA. The Bst polymerase in this kit has good reverse transcriptase activity and can efficiently synthesize cDNA at 60-65℃, so no separate reverse transcription step is required for RNA samples.
• Built-in positive control: A Positive Control is provided to verify the normal performance of the kit. The Positive Control contains positive DNA template and corresponding primers targeting GFP DNA. It is recommended to set up a positive control for each detection.
• Scalable reaction scale: For a standard 20µL reaction system, the small package can perform 25 detections, the medium package 100 detections, and the large package 500 detections.