Lab Book

27.07.10

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

On LB agar plates:

TOP10
MG1655
W3110 hns 93-1
BW25113

On LB agar + kan plates:

BW25113 Δhns::kan
BW25113 ΔtraA::kan

Incubated at 37°C overnight

28.07.10

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.

29.07.10

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

30.07.10

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