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{"date" : "2013-07-27","author" : "fabio","title" : " the blue ligation: the never ending story !","content" : " I first added sap to the o/n digestion( third), then purified and quantified : yield, 32,3 ng/ul.Then again I ligated 006 to 016 folowing the protocol and plated 10 ul in 200 ul of neb10b.In the afternoon I made 6 inocula from the second ligation plates!Today I decided also to take a crack at a fourth ligation with a new strategy: using 006 as my plasmid and 016 as the insert!! I need to digest the two part with different enzymes E and X for 006 and E and S for 016! This time I digested 2400 ng for only 5 ours, not all the night: yields are, 25 ng/ul for 016 and 11,8ng/ul for oo6. Tomorrow I will continue with the ligation number 4.","tags" : " YF1_FixJ - FixK2"}