Transform 1¢X add 100£gL competent cell (bacteria:JM101 or DH5-£\) to the 3 reactions. 2¢X place on ice for 30 min 3¢X 41.5¢XC water bath for 90 sec 4¢X add each tube 1mL LB (without antibiotics) 5¢X 37¢XC water bath for 1 ~2 hours 6¢X spin down and collect about 100£gL LB+bacteria 7¢X spread the bacteria on the Kanamycin plates 8¢X incubate in 37¢XC over night