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Team:Michigan/Nick's Notebook
From 2010.igem.org
Nick's Notebook
10/16/2010
Nick
10-16-10 Psuedomonas Competent Cell Preparation
Full Protocol - Competent Cell Preparation for Quick Transformation of E. coli
Ben Parker started the overnight cultures and assisted in OD measurement and initial centrifugation steps.
A colony of P. putida and P. fluorescence was picked from LB agar plates (10/6/10) and grown up in an overnight culture in 2ml Amp at 30 C.
The overnight culture was added to 100 ml of LB media and grown at 30C for 4 hours.
Protocol was carried out approximately 20 minutes after the following OD600 was recorded.
Blank - water P. putida – 0.587 P. Fluorescence – 0.610
One centrifuge tube containing p. putida culture failed during centrifugation at 5000 rpm. The protocol was altered by reducing the speed to 2500 rpm after this point to reduce the chance of failure. The cells are stored in the 4C fridge in room 1239.
