Isothermal Nucleic Acid Amplification Technology (INAAT) is mainly used in molecular biology to identify and analyze nucleic acids. It is mainly used for DNA, RNA, cellular proteins, ions, molecules, etc. INAAT provides a rapid, sensitive, and accurate diagnosis of genetic and infectious diseases. Its amplified products can be used in biosensors, bioimaging, and biomedicine. In addition, INAAT is used to detect various diseases such as influenza, HIV, tuberculosis, chlamydia, gonorrhea (CT/NG), hepatitis A and hepatitis B, etc. according to the technology, isothermal amplification can be divided into LAMP, RPA, CPA, SDA, RCA, HDA, etc. So what is LAMP? What are the advantages of RT-LAMP?

 

1. What is LAMP?
2. What are the advantages of RT-LAMP？
3. What are the features of the "Cat#13762 RT-LAMP Dye Assay Kit (UDG plus)"?
4. What are the features of the "Cat#13906 RT-LAMP pH Sensitive Dyestuff Kit"?
5. Can it be freeze-dried into freeze-dried beads?
6. Which products can we recommend?

 

1. What is LAMP?

LAMP is currently a widely used technology in the field of isothermal amplification. It uses 4-6 primers that can identify 6 specific regions on the target gene, and relies on the strong strand displacement activity of Bst DNA polymerase. The internal amplification of the target product can reach 10⁹-10¹⁰ times. There are many LAMP detection methods, including the dye method, pH colorimetric method, turbidity method, HNB, calcein, etc. The newly developed fluorescence method and pH detection kits of our company have high sensitivity, good primer applicability, and excellent performance in specificity, tolerance, and stability. Compared with similar products, it has obvious advantages.

 

2. What are the advantages of RT-LAMP？

Easy to operate: short reaction time, low laboratory environment requirements, can be used with microfluidic chips or portable detectors for rapid detection;
Sensitivity: 50 copies/T, 25 μL system;
Good primer applicability: N-2 primer, E gene primer, internal reference primer amplification, etc. have good applicability;
Specificity: 48-well NTC has no peaks at all, and the specificity is strong;
Tolerance: good tolerance to some nucleic acid release agents;
High stability: stable at 37°C for 5 days with good stability, and can withstand repeated freezing and thawing more than 10 times;
Market advantage: The performance is superior to similar domestic and foreign competing products, and the cost performance is high.

 

3. What are the features of the "Cat#13762 RT-LAMP Dye Assay Kit (UDG plus)"?

3.1 High Sensitivity

Figure 1. Sensitivity Test

Red: 50 copies/T, response less than 15 minutes
Blue: 25 copies/T, response less than 15 minutes
The two copy numbers of 50 copies/T and 25 copies/T were tested respectively. The test results showed that within 15 minutes, the results of 50 copies/T performed better, and the detection rate reached 100%.

3.2 Primer Suitability

Figure 2. N5 Locked Nucleic Acid Primer Suitability Detection

Purple: Yeasen     Blue: N Company

Figure 3. N primer suitability assay

Green: Yeasen           Red: N Company

Figure 2, Figure 3: In the specific detection of N5 locked nucleic acid primers and N primers, according to 50 copies/T, the corresponding detection was carried out using Yeasen and N brands. The results showed that the applicability of N5 locked nucleic acid primers was better than that of N brands. The applicability of N primers is the same for the two products.

Figure 4. ACTIN Primer Suitability Detection

Purple: Yeasen          Blue: N Company

15ng of 293DNA was tested for ACTIN primer suitability using Yeasen and N brands. The results showed that Yeasen had excellent applicability.

3.3 Specificity

Figure 5. 48-well NTC-specific assay

48-well NTC detection was used in the specific detection of the product, and no peak was detected, indicating that the specificity was better.

3.4 Tolerance

Figure 6. Nucleic acid releaser tolerance assay

Red: Matrix DEPC H₂O
Green: 50% each of nucleic acid release agent and DEPC H₂O
Blue: 100% Nucleic Acid Release Agent
Using matrix DEPC H₂O, nucleic acid release agent and DEPC H₂O with 50% and 100% nucleic acid release agent were tested respectively. The results show that this product has excellent tolerance to nucleic acid release agents.

3.5 Stability  

Figure 7.Stability testing at the same temperature

Red: 37℃ 0 day  Blue: 37℃ 1 day  Green: 37℃ 3 days  Purple: 37℃ 5 days
0 days at 37°C as a control and stability tests were carried out for 1, 3, and 5 days at the same temperature, indicating that it can still maintain a relatively stable effect at 37°C for 5 days.

Figure 8. -20℃ repeated freeze-thaw stability test

Red: Repeat freeze-thaw 0 times 
Blue: Repeat freeze-thaw 5 times 
Green: Repeat freeze-thaw 10 times

Figure 9. Dry ice repeated freeze-thaw stability test

Red: Repeat freeze-thaw 0 times 
Purple: Repeat freeze-thaw 5 times 
Repeated freeze-thaw stability test at -20°C and dry ice. It was found that the stability of the freeze-thaw was repeated 10 times at -20°C and 5 times on dry ice.

3.6 Comparison of Similar Products

Figure 10. Comparison of the effects of similar products

Red: Yeasen    Blue: Brand N1     Green: Brand N2
Using the primer N gene primer, in the case of 50 copies/T, using Yeasen, brand N1, and brand N2 for product comparison verification, Yeasen products performed well.

  

6. Which products can we recommend?

The products provided by Yeasen are as follows.

Table 1. Product information