Team:Bordeaux/Fourth
From 2012.igem.org
(Created page with "<html lang="en"> <head> <title>Note Book - iGEM Bordeaux 2012</title> <meta charset="utf-8"> <link rel="stylesheet" href="http://cdjemiel.free.fr/igem2012/css/reset.css" type=...")
Newer edit →
Revision as of 13:55, 31 July 2012
Day 15 : 23-07-2012
Today we decided to change some of the biobricks to reduce the number of assemblies
We made a transformation for The first operon biobricks with 2μL DNA.
We also made a Positive control with a well known plasmid and water as a negative control.
We followed the Partregistry protocol as usual.
And Plated 20μL and 200μL of the transformants on LB plates.
We put the plates to grow overnight at 37°C.
Day 11 : 17-07-2012
Day 2 of making new competent cells Read Protocol We stopped incubating our cells at an OD600nm of 0.31 In the end we made 192 tubes of 50μL competent cells.
Day 12 : 18-07-2012
The competence test gave good results with a good amount of colonies on each plates ( ≈ 300 ) and no colonies on the control plate. Today we will transform DNA for every biobrick left. We will use our new competent cells.
We will proceed by heat-shock as described in the partregistry.org protocol Read Protocol
Day 13 : 19-07-2012
Today we picked single colonies and put the to grow in two 5 mL LB + Ampicilin 100μg/mL Except for 3.B who gave no colonies We put the tubes to incubate overnight at 37°C
Day 14 : 20-07-2012
After 13 hours incubating we did a miniprep with the tubes. We got really poor results on the nanodrop for every biobrick (>3ng/μL). As we had really small cells pellets after the first centrifugation we suspect we didn't have enough cells in the beginning.