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Revision as of 09:50, 3 September 2010

Module I, DT and Fan's part:

Measure the concentration of the plasmids purified last night. Double digest to ensure the positive clone.

Double digestion system:

P+E:

 H2O	        6-8μl
 buffer Tango	4μl
 P+E	        4-6μl
 SalI	        2μl
 HindIII	2μl
 Total	        20μl

P+C:

 H2O	        6-11μl		
 buffer Tango	2μl		
 P+C	        4-9μl		
 KpnI	        3μl		
 Total	        20μl	
 37℃ 1h, then	
 + buffer Tango	2.5μl
 + EcoRI	        1μl
   Total	        23.5μl

Run a gel after the digestion is finished.

result

We use the 0.8% agarose gel running first and it is quite un-clear around where 700bp band should be. So we run a 1.2% gel but it seemed that there is no positive result among the 12 clone.