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EPF-Lausanne/11 August 2008
From 2008.igem.org
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Molecular Biology
We try by different way to assemble biobrick
We(Vincent) do a mistake with kanamycin stock, we put 1000 time less the dose.
We start cell culture of all interestings biobrick to do tomorrow fresh mini-prep. Because the mini-prep that we use are not good(kanamycin 1000 time less concentrate).
E1010 and F1610
We try last week to assemble two big parts because it is easy to purify it. We do ligation and transformation last week. The result was great all of the ten plates work. Monday, we decide to do a mini prep of these cells to purify DNA. Then we want to check the plasmid so we digest a small quantity of them with EcoRI and SpeI to prepare it to do a gel tomorrow.
by Gel
We try again to purify insert and vector by Gel.
We try to assemble :
R0040/B0034
R0071/B0034
and I1466/I1433
We successful have isolate from Gel : R0040, R0071 and B0034
We try to change strategy of isolate I1466 and I14033
I1466/I14033
I14033 is small. We let it in the vector. I1466 is big we use it to a insert. We cut with SpeI and PstI. We digest the plasmid and we are going to do tomorrow ligation.
Without gel, use PCR with biobrick primer
We will start tomorrow with new mini-prep to amplify DNA by PCR, by this way we don't need to use Gel purification(that is hard with small fragment).
http://openwetware.org/wiki/Knight:Annealing_and_primer_extension_with_Klenow_polymerase
