Team:Tsinghua/Notebook/12 August 2010

From 2010.igem.org

(Difference between revisions)
(== result ==)
(Module I, DT and Fan's part:)
 
(One intermediate revision not shown)
Line 1: Line 1:
-
== Module I, DT and Fan's part: ==
+
== 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.
Line 22: Line 22:
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.
-
 
-
 
== result ==
== result ==
-
[[Image:10-8-12]]
+
[[Image: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.
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.

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

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.