Team:Tsinghua/Notebook/26 July 2010

From 2010.igem.org

(Difference between revisions)
(New page: == Module II, Danyang's part: == To test the effectivity of newly-synthetized, PCR assay to amplify mCherry, Kan and eGFP, Chlr is conducted. PCR system (FastPFU): H2O ...)
(2c)
 
(One intermediate revision not shown)
Line 1: Line 1:
-
== Module II, Danyang's part: ==
+
== Module I, Group 2c ==
-
To test the effectivity of newly-synthetized, PCR assay to amplify mCherry, Kan and eGFP, Chlr is conducted.
+
To test the effectivity of newly-synthetized primer, PCR assay to amplify mCherry, Kan and eGFP, Chlr is conducted.
PCR system (FastPFU):
PCR system (FastPFU):
Line 16: Line 16:
-
PCR program
+
PCR program----
 +
 
   1.95℃                       1min
   1.95℃                       1min
Line 27: Line 28:
   5.72℃                      5min
   5.72℃                      5min
-
----
 

Latest revision as of 07:27, 25 October 2010

Module I, Group 2c

To test the effectivity of newly-synthetized primer, PCR assay to amplify mCherry, Kan and eGFP, Chlr is conducted.

PCR system (FastPFU):

 H2O	                       31.5μl
 5×buffer	               10μl
 dNTP	                       5μl
 primer1	               1μl
 primer2	               1μl
 template	               1μl
 FastPFU	               0.5μl
 Total	                       50μl


PCR program----


 1.95℃	                       1min
 2.95℃ 	               20s
 3.53℃for eGFP  	        
   51℃for mCherry	       20s
   51℃for Kan&Chlr	       
 4.72℃	                       30s
 30 cycles from 2 to 4
 5.72℃                       5min


Result for PCR: After agarose gel is run, no specific bands are observed.


Conclusion: reaction condition for PCR is not ideal enough, thus we shall change the condition and try this again.