IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/06/27/Zhangs

Miniprep CI plasmid Some improvements: 1.the EB be put in 65°C water bathing before elution; 2.50 uL EB to elute; 3.after PW liquid, centrifuge 10 min before the next step; 4.the rpm increased to 13000 per min A260/A280 = 1.92 A260 = 0.120  CI = 300ng.uL

Previous double digestion protocol seemed to be lack of efficiency, so we tried new protocol today. New double digestion system: For front insert:

For front vector:

Electrophoresis to test Samples: 10μL digestion system+2μL DNA Dye Control: 5μL plasmid+2μL DNA Dye Marker: 10μL DL2000 plus+2μL DNA Dye React for 10 minutes Result (from left to right): Marker, CI control, CI, 1-23L control, 1-23L, 1-4H control, 1-4H

Gel extraction Extract CI insert, 1-4H vector and 1-23L vector