Team:Tsinghua/Notebook/9 August 2010

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(Difference between revisions)
(New page: == Module I, DT and Fan's part: == Measure the concentration of the plasmids purified last night. Double digest to ensure the positive clone. Double digestion system: P+E: H2O ...)
(Module I, DT and Fan's part:)
 
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== Module I, DT and Fan's part: ==
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== Module I, group 2(b) ==
Measure the concentration of the plasmids purified last night. Double digest to ensure the positive clone.
Measure the concentration of the plasmids purified last night. Double digest to ensure the positive clone.
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   Total         20μl
   Total         20μl
   37℃ 1h, then
   37℃ 1h, then
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   + buffer Tango 2.5μl
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   + buffer Tango     2.5μl
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   + EcoRI         1μl
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   + EcoRI     1μl
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     Total         23.5μl
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     Total     23.5μl
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Run a gel after the digestion is finished.
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Run a gel after the digestion is finished.
== result ==
== result ==
We use the 0.8% agarose gel running first and it is quite un-clear around where 700bp band should be. So we run a 1.2% gel but it seemed that there is no positive result among the 12 clone.
We use the 0.8% agarose gel running first and it is quite un-clear around where 700bp band should be. So we run a 1.2% gel but it seemed that there is no positive result among the 12 clone.

Latest revision as of 06:54, 25 October 2010

Module I, group 2(b)

Measure the concentration of the plasmids purified last night. Double digest to ensure the positive clone.

Double digestion system:

P+E:

 H2O	        6-8μl
 buffer Tango	4μl
 P+E	        4-6μl
 SalI	        2μl
 HindIII	2μl
 Total	        20μl

P+C:

 H2O	        6-11μl		
 buffer Tango	2μl		
 P+C	        4-9μl		
 KpnI	        3μl		
 Total	        20μl	
 37℃ 1h, then	
 + buffer Tango	    2.5μl
 + EcoRI	    1μl
   Total	    23.5μl

Run a gel after the digestion is finished.

result

We use the 0.8% agarose gel running first and it is quite un-clear around where 700bp band should be. So we run a 1.2% gel but it seemed that there is no positive result among the 12 clone.