Establishment of mink heart identification method based on mitochondrial cytochrome b gene and development of its detection kit

摘要

In this study, the mink heart mitochondrial DNA was used as the target gene to design the specific primers of mink heart mtDNA Cytb, the extraction and detection reagents of mink heart DNA were developed using DNA fingerprinting technique, and the specificity, reproducibility, and stability of the reagents were investigated. Molecular cloning and sequencing technique were used to clone the standard substance of mink heart DNA detection, then a DNA fingerprint detection method of mink heart and the quality standard for mink heart were established, and a DNA detection kit of mink heart was developed. The results showed that the structure of DNA extracted from mink heart by self-developed reagents was complete, and both the concentration and purity ofDNA were high. A specific amplification band of the original mink samples was found at 337 bp. The sequence of mink heart DNA was consistent with that of mink heart mtDNA specific fingerprint region. The mink heart DNA fingerprint identification method established, in this study, is accurate and reliable, and the procedure of the developed DNA kit is easy, the results obtained using this kit is stable and the method is suitable for popularization and application.