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Team:Cambridge/LabBook/Week3
From 2010.igem.org
(Difference between revisions)
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Incubated at 37°C overnight | Incubated at 37°C overnight | ||
==28.07.10== | ==28.07.10== | ||
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all strains grew with individual colonies. | all strains grew with individual colonies. | ||
===E2.Set up overnight culture of TOP10 (Emily & Anja)=== | ===E2.Set up overnight culture of TOP10 (Emily & Anja)=== | ||
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| 0.530 | | 0.530 | ||
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| + | ===Preparation CCMB80 Buffer (Volume500ml)=== | ||
| + | Arrows label dilutions to the indicated concentration | ||
| + | KOAC 1M ---> 10mM 5ml | ||
| + | CaCl<sub>2</sub>.2H<sub>2</sub>0 1M ---> 80mM 40ml | ||
| + | MnCl<sub>2</sub>.4H<sub>2</sub>0 1M ---> 20mM 10ml | ||
| + | MnCl<sub>2</sub>.6H<sub>2</sub>0 1M ---> 10mM 5ml | ||
| + | glycerol --->10% | ||
| + | sterile H<sub>2</sub>0 390ml | ||
| + | prepare in flow hood | ||
| + | sterile filter | ||
| + | store at 4°C | ||
Revision as of 13:17, 6 August 2010
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
