After plasmid or cosmid DNA purification using the endofree plasmid maxi kit

product: 3ug/ul, 50ul 

plasmid 10kb

NOT1 digestion: 

 system        total  200ul

                   10*buffer o 20ul

                   DNA 40ul

                   NOT1 25ul (10unit enzyme needed for 1 ug DNA, we use 120ug DNA)

                   ddH2O 115ul

37^& for 3hr.

0.8% agarose electrophoresis to detect the result.   OK

 

After that , we purify the digestion product. using QIAquick PCR Purification Kit

 

points before starting: add 100% ethanol to buffer PE.

PROCEDURE:     see next chapter 2nd April. 2009