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From 2010.igem.org

Contents 1 Lab Book 1.1 27.07.10 1.1.1 1. Streaking out of bacterial cultures (Peter & Anja) 1.2 28.07.10 1.2.1 E2.Set up overnight culture of TOP10 (Emily & Anja) 1.3 29.07.10 1.3.1 Preparation CCMB80 Buffer (Volume500ml)

Lab Book

27.07.10

1. 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.

E2.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)
12:20	-0.026	-0.032
2:30	0.437	0.530

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