Dual-Enzyme CRISPR-Based Nucle

This product utilizes a chromatographic double-antibody sandwich method for the detection of enzyme cleavage products from Cas12 and Cas13. Following the amplification of the target gene via LAMP, RAA/RPA, and PCR, Cas12 and Cas13 enzymes are employed simultaneously to cleave the amplified product. Detection of cleavage products becomes feasible upon modification of DNA signal probe A corresponding to Cas12 enzyme cleavage, or RNA signal probe B corresponding to Cas13 enzyme cleavage.

Cat. No.: DCLFS-50 (for 50T)

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Description

This product utilizes a chromatographic double-antibody sandwich method for the detection of enzyme cleavage products from Cas12 and Cas13. Following the amplification of the target gene via LAMP, RAA/RPA, and PCR, Cas12 and Cas13 enzymes are employed simultaneously to cleave the amplified product. Detection of cleavage products becomes feasible upon modification of DNA signal probe A corresponding to Cas12 enzyme cleavage, or RNA signal probe B corresponding to Cas13 enzyme cleavage, as follows:

1. Probe A is biotin-modified at one end and fluorescein isothiocyanate (FITC) or 6-carboxyfluorescein (6-FAM) modified at the other end. Probe B is biotin-modified at one end and digoxin (Dig) modified at the other end.

2. Alternatively, probe A is biotin-modified at one end and digoxin (Dig) modified at the other end, while probe B is biotin-modified at one end and fluorescein isothiocyanate (FITC) or 6-carboxyfluorescein (6-FAM) modified at the other end.

Instruction

1. Retrieve the appropriate number of test strips corresponding to the number of test samples and mark them on the absorbent pad. Each test strip is intended for a one-time single sample test. If the volume of the amplification product falls between 50-100µL, the nucleic acid product can be directly detected within the 200µL PCR reaction tube. However, if the amplification product volume is less than 50µL, ultrapure water must be added to the PCR tube to adjust the volume to 50µL. Detection can only proceed after thorough mixing by pipetting.

2. After completing the PCR, RPA, or RAA amplification, add the products to the CRISPR system. Open the PCR reaction tube and carefully insert the binding pad end (arrow end) of the test strip into the PCR reaction tube. Ensure that the liquid level does not exceed the top of the binding pad. Once the reading area becomes fully saturated (approximately 1-2 minutes, with potential variations due to external temperature conditions, such as slower water absorption in colder environments like winter, prolonging the saturation time), and after the color of the quality control line (C line) appears, remove the test strip. Interpret the test result directly based on the color development of the test strip.

3. Ensure to read the results within 10 minutes after the appearance of the quality control line (C line) color. Any interpretation after 10 minutes is considered invalid.

4. Record the test outcomes, securely seal, and dispose of the test strips in a safe location.

Storage