Team:Tsinghua/Notebook/12 August 2010


Module I, group 2(b)

Use the K-up and K-do to detect the P+K+C. Meanwhile use the digestion protocol to ensure the result.

Double digestion system:


 H2O	         5.5-10.5μl
 buffer BamHI	 2μl
 P+E+M	         5-10μl
 SalI	         1.5μl
 BamHI	         1μl
 Total	         20μl


 H2O	         0-10.5μl
 buffer BamHI	 2μl
 P+K+C	         5-16μl
 KpnI	         1.5μl
 BamHI	         1μl
 Total	         20μl

Run a gel. Order: P+E+M 1-6 double digestion, Marker, P+K+C 1-6 double digestion, P+K+C PCR.



P+E+M② is positive. It seems that PKC 1-6 are positive. But it still remains unclear whether the PCR bands are so weak and why the digestion result seemed negative.