then using gel transparent spectrum method to analyze gel molecule content; using NaH2PO4, Na2SO4 and dodecyl sodium sulfate as flow phase; the column analysis adopts protein molecule column; the tester adopts ultraviolet tester and difference tester with wavelength of 190 to 230nm and temperature ...
Cells were harvested, resuspended in lysis buffer (50 mM NaH2PO4, 300 mM NaCl, 10 mM imidazole, pH 8.0) and lysed by sonication at 4°C. Soluble rEhRAD51 was purified near to homogeneity under denaturing and native conditions through Ni2+-NTA affinity chromatography according to the ...
After 16 h-growth at 30 °C, cells were harvested, resuspended in lysis buffer (50 mM NaH2PO4 pH 8.0, 300 mM NaCl, 10 mM Imidazole) and lysed by sonication. Cell lysate was clarified by centrifugation and incubated with an immobilized metal-affinity matrix (Ni-NTA Agarose, ABT ...
The cells were washed with ice-cold PBS (2.67 mM KCl, 1.5 mM KH2PO4, 137 mM NaCl and 8.1 mM NaH2PO4, pH 7.4) and collected by scraping. The cells were pelleted by centrifugation at 300g for 3 min. The cell pellets were flash-frozen in liquid N2 and stored at −...
Duplicate nitrocellulose membrane plaque lifts were washed with 2 × SSPE (0.2 M NaH2PO4, pH 7.4, 3 M NaCl, 20 mM EDTA), 0.5% SDS during 1 h at 55°C and subjected to prehybridization and hybridization for 4 and 12 h, respectively, at 50°C in 5 × SSPE, containing 2.5× Denhardt...
RNA was preincubated with 3.5 μl of 2× binding buffer (20 mM Tris·HCl, pH 8.0; 1 mM DTT; 1 mM MgCl2; 20 mM KCl; and 10 mM Na2HPO4–NaH2PO4, pH 8.0) for 30 min at 37°C (modified from Maki et al., 2008) and translation was performed using PURExpress In...
Each culture was pelleted, resuspended in 25 ml LEW buffer (50 mM NaH2PO4, 300 mM NaCl, pH 8.0) and incubated at 4°C for 30 min with 1 mg ml−1 lysozyme. The samples were then lysed by sonication and subsequently centrifuged for 30 min at 5000 g. The resulting...
Bacteria were harvested and resus- pended in denaturing buffer (100 mM NaH2PO4, 10 mM Tris Cl and 8 M Urea). The 6xHis tagged-recombinant pro- tein was then purified by Ni2+ nitrilotriacetic acid (Ni- NTA) affinity chromatography under denaturing condi- tions using QIAexpressionist™ ...
clamped homogeneous electric fields (CHEF) PFGE electrode array constructed as described by Chu et al28 High molecular weight DNA was digested, fractionated by CHEF, and then transferred to a nylon membrane (Nytran; Schleicher and Schuell, Inc, Keene, NH) in 10× SSPE (NaCl, NaH2PO4, and ...
Cells were harvested, resuspended in lysis buffer (50 mM NaH2PO4, 300 mM NaCl, 10 mM imidazole, pH 8.0) and lysed by sonication at 4°C. Soluble rEhRAD51 was purified near to homogeneity under denaturing and native conditions through Ni2+-NTA affinity chromatography according to the ...