Team:Cambridge/LabBook/Week3

From 2010.igem.org

(Difference between revisions)
Line 1: Line 1:
=Lab Book=
=Lab Book=
==27.07.10==
==27.07.10==
-
===1. Streaking out of bacterial cultures (Peter & Anja)===
+
===1. Experiment: Streaking out of bacterial cultures (Peter & Anja)===
On LB agar plates:
On LB agar plates:
  TOP10
  TOP10
Line 13: Line 13:
==28.07.10==
==28.07.10==
all strains grew with individual colonies.  
all strains grew with individual colonies.  
-
===E2.Set up overnight culture of TOP10 (Emily & Anja)===
+
===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 single bacterial colony (ATP10) in 12ml LB, incubated at 37°C with shaking (180 rpm) overnight.
==29.07.10==
==29.07.10==
Line 25: Line 25:
! OD<sub>600</sub> (2)
! OD<sub>600</sub> (2)
|-
|-
-
| 12:20
+
| 12:20pm
| -0.026
| -0.026
| -0.032
| -0.032
|-
|-
-
| 2:30
+
| 2:30pm
| 0.437
| 0.437
| 0.530
| 0.530
|}
|}
-
===Preparation CCMB80 Buffer (Volume500ml)===
+
===3. Preparation: CCMB80 Buffer (Volume500ml)===
Arrows label dilutions to the indicated concentration  
Arrows label dilutions to the indicated concentration  
  KOAC 1M ---> 10mM 5ml
  KOAC 1M ---> 10mM 5ml
Line 45: Line 45:
  sterile filter
  sterile filter
  store at 4°C
  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
 +
 +
OD<sub>600</sub> measurements:
 +
{| class="wikitable"
 +
|-
 +
! Time
 +
! OD<sub>600</sub> (teaching lab)
 +
! OD<sub>600</sub> (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
 +
|}

Revision as of 13:29, 6 August 2010

Contents

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

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
Retrieved from "http://2010.igem.org/Team:Cambridge/LabBook/Week3"