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From 2013.igem.org

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Revision as of 13:46, 19 August 2013

{ "date" : "2013-08-01", "author" : "fabio", "title" : " the blue ligation: the never ending story !", "content" : " today I made the Onetaq pcr on my positive ligation miniprep (same program and sample composition that I used last time!! On the gel below we can see 016 digested and my positive ligation pcr amplification.

After this success I started the ligation of this device to r0010, already digested with proper enzymes: E and S; I digested with E and X for 30 minutes even 50 ng O16+006 ( yield is 161,9 ), then deactivated enzymes at 80 degrees, added Sap, and ligated: I prepared control, 1:1, 1:2, 1:3 and 1:4 using the ligation protocol, and incubating for 30 minutes, then I deactivated at 80 degrees. After that I transformed 10 ul of the ligations in neb10b. ", "tags" : " YF1_FixJ - FixK2- lacI" }