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Team:Tsinghua/Notebook/12 August 2010
From 2010.igem.org
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| - | == Module I, | + | == 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. | 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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Run a gel. Order: P+E+M 1-6 double digestion, Marker, P+K+C 1-6 double digestion, P+K+C PCR. | Run a gel. Order: P+E+M 1-6 double digestion, Marker, P+K+C 1-6 double digestion, P+K+C PCR. | ||
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== result == | == result == | ||
Latest revision as of 06:54, 25 October 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:
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
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.
