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From 2013.igem.org

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Revision as of 13:31, 19 August 2013

{ "date" : "2013-07-05", "author" : "emil", "title" : " Purification of the second run of Inocula of ligation and re-ligation of the digested products ", "content" : " I added 1µl of DPN1 to the insert and 1µl of SAP to the backbones.Then I have purified the inocula following the miniprep protocol.Afterwards I quantified them toghether with the results of the purification of the digestion products(performed following the purification protocol ) with the following results:

Then I digested 800 ng of each sample with EcoR1 HF and Pst1 following the digestion protocol with the following results:As we can see no one of the wells shows the right insert in the lower band(there is always the previous insert the RFP + Lac promoter ~ 1200 bp).At the same time I did the ligation of K823024+E0840 and K823026+E0840 previously digested following the ligation protocol .
N.B. Remember always to spin the ligation buffer before using
After that I plated the product of the ligation as done before(1:1,1:3,ctrl for each sample) on Ampicillin added LB.", "tags" : "K832024-K823026-E0840" }