Team:Cambridge/LabBook/Week3

From 2010.igem.org

Revision as of 13:17, 6 August 2010 by Willh (Talk | contribs)

Contents

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