Note Book - iGEM Bordeaux 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 16 : 24-07-2012

Presence of colonies on both the positive and the negative control , we suspect a resistance from the competant cells or a bad selection with the plates.
We ran a test to check the cells and the plates by puting to incubate overnight at 37°C.
one ampicilin plate + ampicilin extempo + competent cells , one ampicilin plate + competent cells and one empty plate.

Day 17 : 25-07-2012

The plates were responsible of the non selection of the transformants.
We made a new stock of LB plates with Ampicilin , Chlorenphenicol or Kanamicin.
We transformed the biobricks from operon 1 an 3 again but plated them on our new stock.
Except for I.A put into pre-culture of 2mL LB and I.B as we used the transformant from Day 15.

Day 18 : 26-07-2012

Today we did a mini prep for each 5 pre culture tubes of I.A
We transformed the rest of the biobricks , IC/IIB/IIC(2)/IID/IIIB/IVA/IVC
We put five colonies of ID/IE/IIIA/IIIC/IIID/IIIE/5A into a 2mL LB pre-culture

Day 19 : 27-07-2012

Today we did a miniprep with the colonies put into pre-culture overnight.
We put five colonies of IB/IC/IIB/IIC(2)/IID/IIIB/IVA/IVC into a 2mL LB pre-culture.