RNA extraction from yak milk for amplification of specific protein encoding genes of mammary gland
In order to establish a method for extracting RNA from milk and amplifying genes,this study carried out preliminary i-solation of exosomes from the mixed milk of four Maiwa yaks by ultracentrifugation,followed by total RNA extraction by TRIzol method,and the coding regions of three specific genes in mammary epithelial cells were amplified by PCR after reverse tran-scription and directly sequenced.The results showed that the RNA extracted from the milk was of good quality and could be used to amplify the complete coding regions of κ-casein gene and β-lactoglobulin gene,with lengths of 705 bp and 636 bp,re-spectively,and the products could be directly sequenced,but failed to amplify the lactoferrin gene.In addition,this experiment preliminarily showed that the number of PCR cycles using RNA extracted from yak milk should be between 35 and 45 cycles.In this study,a new method for RNA extraction based on exosomes in yak milk was established,and the extracted RNA could be used for gene amplification and sequencing.
NETL
NSTL
万方数据
维普
