"

From 2010.igem.org

Module I, group 2(b)

Measure the concentration of the PCR products.

Digest the four fragments with follow system at 37℃ for 10 hours.

Digestion system:

eGFP:

 H2O	        23.5μl
 10×buffer	5μl
 eGFP	        20μl
 HindIII	1.5μl
 Total	        50μl

mCherry:

 H2O	        11.5μl
 10×buffer	5μl
 mCherry	32μl
 HindIII	1.5μl
 Total	        50μl

Kan:

 H2O	        31μl
 10×buffer	5μl
 Kan	        12.5μl
 KpnI	        1.5μl
 Total	        50μl

Chlr:

 H2O	        31.5μl
 10×buffer	5μl
 Chlr	        12μl
 KpnI	        1.5μl
 Total	        50μl

Productions purify and measure the concentration.

 Gene	    Concentration(μg/ml)	OD 260/280
 eGFP	        38.4	                  1.66
 mCherry	28.8	                  1.86
 Kan	        43.7	                  2.11
 Chlr	        31.9	                  1.77