EPF-Lausanne/11 August 2008

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(Difference between revisions)
(Molecular Biology)
(Molecular Biology)
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We try by different way to assemble biobrick
We try by different way to assemble biobrick
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We(Vincent) do a mistake  
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We(Vincent) do a mistake with kanamycin stock, we put 1000 time less the dose.
==E1010 and F1610==
==E1010 and F1610==
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==Without gel, use PCR with biobrick primer==
==Without gel, use PCR with biobrick primer==
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We start cell culture of all interestings biobrick to do tomorrow fresh mini-prep. Because the mini-prep that we use are not good.
+
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).
http://openwetware.org/wiki/Knight:Annealing_and_primer_extension_with_Klenow_polymerase
http://openwetware.org/wiki/Knight:Annealing_and_primer_extension_with_Klenow_polymerase

Revision as of 06:51, 13 August 2008

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Contents

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.

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 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).

http://openwetware.org/wiki/Knight:Annealing_and_primer_extension_with_Klenow_polymerase