Objective:

2

To establish a rapid on-site method for identifying Chinese medical material recombinase-mediated amplification (RAA) technology for the use of identifying medicinal Bubali Cornu from yak horn.

Method:

2

Based on the differences of mitochondrial genome sequences between Bubali Cornu and adulterants

the specific RAA primer (SNJ-1.F

SNJ-1.R) and fluorescence probe SNJ-1.probe were designed by variation sites. Alkaline lysis method was used to extract DNA from milled samples

and optimize RAA reaction system. The incubation was made at 37 ℃ for 15-20 min

the reaction results were monitored through gel electrophoresis and a mobile fluorescence amplification instrument. The RAA identification result was compared with COI DNA sequencing.

Result:

2

After incubation at 37 ℃ for 20 min

about 140 bp of bright and simple bands was amplified from DNA templates of

Bubalus bubalis

whereas

Bos mutus

were negative. By the Real-time fluorescent RAA identify method

all reactions in DNAs from Bubali Cornu samples were amplified from 6.21 to 8.37 min

whereas DNAs from yak samples were amplified after 10.08 min

COI sequencing results conformed to the Real-time fluorescent RAA identification.

Conclusion:

2

Specific RAA could rapidly identify Bubali Cornu in 20 minutes

and thus be applied in medical material and its products. This method is simple

rapid and sophisticated instrument-free

with promises in on-site identification of traditional Chinese medicine.