赛默飞世尔Thermo Fisher X12 BOTTLE

原创：western protocol

albumin (Fraction V) Make up in PBST and sterile filter. Keep at 4°C to prevent bacterial contamination. 10xPBST 1L PH=7.4 Nacl 80g KCl 2g Na2HPO4 144g KH2PO4 0.24g Tween20 3ml 10xSecondary washing solution 1L PH=7.5 150mM NaCl 8.766g 50mM

超级感受态细胞的制备

, or equivalent). Adjust the pH of the solution to 6.7 with 5 M KOH, and then add pure H2 O to bring the final volume to 100 ml. Sterilize the solution by filtration through a disposable prerinsed Nalgene filter (0.45-mm pore size). Divide into aliquots and store

Methods for DNA isolation

or cosmid DNA of interest into a 12 X 75 mm Falcon tube containing 2 ml of LB media supplemented with the appropriate antibiotic (typically ampicillin at 100 ug/ml) and incubate at 37deg C 8-10 hours with shaking at 250 rpm. Transfer the culture