Tris saturated phenol chloroform isoamyl alcohol

Cephradine capsules (batch number: 151101) were purchased from Suzhou Chung-Hwa Chemical & Pharmaceutical Industrial Co. Ltd., and gentamicin sulfate (batch number: 5150307) was purchased from Yichang Pharmaceutical Industrial Co. Ltd. The two antibiotics were then prepared into a mixture at a concentration of 62.5 g·L^-1[26 (link)]. The DNA extraction reagents (such as Proteinase K, lysozyme, Tris-saturated phenol-chloroform-isoamyl alcohol (25:24:1) and TE buffer) were purchased from Beijing Dingguo Changsheng Biotechnology Co. Ltd., and prepared in the lab (such as 10% SDS, chloroform-isoamyl alcohol (24:1), 5 mol·L^-1 NaCl, 0.1 mol·L^-1 PBS and CTAB/NaCl). D. hansenii, which was provided by the laboratory and shaken at 28°C for 36 hours after being inoculated into liquid Potato Sucrose medium in a 300 mL erlenmeyer flask. The cells were then gathered by centrifugation at 2000×g for 4 minutes after washed 1∼2 times repeatedly with sterile stroke-physiological saline solution. The above cells were diluted to 10¹⁰ mL^-1 with sterile storke-physiological saline solution eventually after being counted by hemocytometer, and stored at 4°C for subsequent experiments[27 (link)].

Cephradine capsules (product batch number: 110804) and gentamicin sulfate injection (product batch number: 5120106) were purchased from Suzhou Chung-Hwa Chemical & Pharmaceutical Industrial Co., Ltd. and Yichang Renfu Pharmaceutical Co., Ltd., respectively. An antibiotic mixture was prepared at a concentration of 62.5 g L⁻¹ with sterile saline according to the ratio of 1 : 2 (i.e., 6 gentamicin (2 mL/branch) + 3 cephalosporins (0.25 g/grain)) and stored at 4°C [16 (link), 17 ]. Protease K, Tris-saturated phenol-chloroform-isoamyl alcohol (25 : 24 : 1), lysozyme, and TE buffer were purchased from Beijing Ding Guo Biotechnology Co., Ltd. Other reagents were prepared in the lab.