Loop-mediated isothermal amplification design four primers for six regions on the target gene, and use stranded DNA polymerase to perform the amplification reaction at a constant temperature (60-65℃), which can achieve 109-1010 within 15-60 minutes The multi-fold amplification is simple, rapid and specific.


Types of section:
LAMP-PCR
RT-LAMP-PCR

Fluorescent dyes:
SYBR Green is a DNA binding dye that can be incorporated into double-stranded DNA non-specifically. In the free state, it does not emit fluorescence, but once bound to double-stranded DNA, it can emit fluorescence. During the PCR reaction, as the cycle increases, the amount of double-stranded DNA increases, and the corresponding SYBR Green fluorescence value increases, thereby indicating the running process of the PCR reaction.


UNG enzyme anti-pollution:
UNG enzyme can selectively hydrolyze and break uracil glycosidic bonds in double-stranded or single-stranded DNA containing dU. The most important pollutant in PCR experiments is PCR products. To prevent contamination, dUTP is used to replace dTTP in conventional PCR reaction systems. Generate DNA amplification products containing dUTP, add a 50°C heat preservation step before the next PCR, UNG enzyme can degrade the existing U-DNA contaminants in the reaction system, thereby ensuring the specificity and accuracy of the amplification results , And then the UNG enzyme is inactivated at the denaturation temperature.

Advantage:
1. Fast and efficient. There is no need for thermal denaturation of double-stranded DNA in advance, avoiding time loss caused by temperature cycling.
2. High sensitivity, several orders of magnitude higher than PCR.
3. High specificity. There are 4 specific primers designed for 6 regions of the target sequence. If one of the 6 regions does not match the primer, nucleic acid cannot be amplified.
4. Simple operation, no need for expensive instruments, simple requirements and strong operability.

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