Revision as of 13:29, 6 August 2010 by Willh (Talk | contribs)


Lab Book


1. Experiment: Streaking out of bacterial cultures (Peter & Anja)

On LB agar plates:

W3110 hns 93-1

On LB agar + kan plates:

BW25113 Δhns::kan
BW25113 ΔtraA::kan

Incubated at 37°C overnight


all strains grew with individual colonies.

2. Experiment: Set up overnight culture of TOP10 (Emily & Anja)

Inoculated single bacterial colony (ATP10) in 12ml LB, incubated at 37°C with shaking (180 rpm) overnight.


Inoculated 400ml LB with 5ml TOP10 overnight culture, incubate at 37°C with shaking (180rpm) put on at 11:40am

OD600 measurements:

Time OD600 (1) OD600 (2)
12:20pm -0.026 -0.032
2:30pm 0.437 0.530

3. Preparation: CCMB80 Buffer (Volume500ml)

Arrows label dilutions to the indicated concentration

KOAC 1M ---> 10mM 5ml
CaCl2.2H20 1M ---> 80mM 40ml
MnCl2.4H20 1M ---> 20mM 10ml
MnCl2.6H20 1M ---> 10mM 5ml
glycerol --->10%
sterile H20 390ml 
prepare in flow hood
sterile filter
store at 4°C

4. Experiment: Set up overnight cultures of TOP10 (Will & Anja)

Inoculated single bacterial (TOP10) colony in 5ml SOB, incubated at rtp with shaking overnight


5.Experiment: Preparing chemicall competent cells (Emily, Bill & Anja)

(followed protocol for 'TOP10 chemically competent cells' from OpenWetWare)

Inoculated 800ml SOB with 3ml of TOP10 overnight culture (grown from single colony), incubated at 37°C with shaking (180rpm)

Put on at 9.45am

OD600 measurements:

Time OD600 (teaching lab) OD600 (Jim's lab)
10:50am 0.008 -0.002
11:30am 0.013 0.000
12:00pm 0.023 0.024
12:35pm 0.032 0.034
01:00pm 0.040 0.046
01:32pm 0.052 0.063
02:00pm 0.073 0.090
02:35pm 0.106 0.140
03:00pm 0.137 0.174
03:25pm 0.179 0.230
03:38pm 0.203 0.256
03:55pm 0.225 0.313