Team:Tsinghua/Notebook/24 September 2010

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(Difference between revisions)
(New page: == 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: P+E+M: H2O 5.5-10.5...)
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== Module I, group 2(b) ==
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== Module , group 4 ==
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Use the K-up and K-do to detect the P+K+C. Meanwhile use the digestion protocol to ensure the result.
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After getting the primer,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:  
Double digestion system:  

Revision as of 02:19, 28 October 2010

Module Ⅱ, group 4

After getting the primer,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:

P+E+M:

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

P+K+C:

 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.

result

10-8-12.jpg

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.