[Application of allele-specific primer in the identificatin of Hedyotis diffusa].

OBJECTIVE

To develop a convenient and effective method for the identification of Hedyotis diffusa.

METHODS

DNA templates were extracted from H. diffusa and its adulterants samples on commercial markets. And DNA fragments of rDNA ITS-2 regions were amplified and sequenced subsequently. ITS-2 sequences of all samples were aligned. The allele-specific primer was designed for distinguishing H. diffusa.

RESULTS

The nucleotide difference between H. diffusa and 8 other species is obvious in the ITS-2 region. In addition, the allele-specific primers were employed to amplify the DNA from H. diffusa and 8 other species. The result indicated that a 392 bp DNA fragment was amplified from H. diffusa, whereas no any fragment was amplified from 8 other species under the same reaction condition.

CONCLUSION

The primers designed in the present study were highly specific for H. diffusa. They could be used as key components in the H. diffusa identification kit.