Team:Freiburg/Notebook/26 August

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<a href="https://2011.igem.org/Team:Freiburg/Notebook/25_August">Previous entry</a>
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<a href="https://2011.igem.org/Team:Freiburg/Notebook"> 26 August </a>
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==Meeting==
==Meeting==
Line 47: Line 61:
==<span style="color:green;">green light receptor</span>==
==<span style="color:green;">green light receptor</span>==
-
===NAME OF YOUR EXPERIMENT===
+
===repeating quickchange PCR===
-
'''Investigators:NAME'''
+
'''Investigators:Julia'''
 +
<br/>
 +
repeating PCR with fresh DNA template.
 +
<br/>
 +
digest with DpnI,added directly to the PCR tube.<br/> After NEB the DPNI has also a good activity in Phusion HF buffer.<br/>
 +
==<span style="color:blue;">blue light receptor</span>==
 +
===Ligation===
 +
'''Investigators: Sophie'''<br/>
 +
Date: 25.08.11<br/>
 +
continue from experiment: Digestion; 
 +
Investigator: Sandra<br/> 
 +
Project Name: Blue light receptor<br/>
 +
all samples stored in Blue light box
-
==<span style="color:blue;">blue light receptor</span>==
 
-
===NAME OF YOUR EXPERIMENT===
+
===Transformation===
-
'''Investigators:NAME'''
+
'''Investigators: Sophie'''<br/>
 +
Date: 26.08.11<br/>
 +
Continue from                                  Date: 26.08.11        Name: Sophie               
 +
Experiment Ligation <br/> 
 +
Project Name: Blue light receptor<br/>
 +
Procedure
 +
<br/>
 +
1. take cells from -80°C freezer and put them on ice! (every eppi contains about 400 μl cells)
 +
2. thaw cells on ice 20 minutes
 +
3. pipette 50 μl cells and 2 μl DNA into eppi still on ice!
 +
4. Incubate for 30 minutes on ice
 +
5. Heat at 42°C for 60 sec
 +
6. Incubate on ice for 5 minutes
 +
7. Add 200 μl LB Broth
 +
8. Incubate for 2 hours at 37°C (cells with lysis cassette at 30°C!!)
 +
9. Plate 50 μl and 200μl on two different LB/Agar plates with appropriate antibiotic resistance
 +
 +
<br/>
 +
Documentation:
 +
Why are you doing this experiment? Name of the sample? Where are they stored? Name the vector with inserts, antibiotika resistance etc.
 +
Why? The last cloning was done with a NOT-PCR-Product which might make problems because it does not have enough bases after the restriciton sites.<br/>
 +
 +
Name of the samples: Not 1:1, Not 1:3, nOt 1:1 nOt 1:3, noT 1:1, noT 1: 3<br/>
 +
stored in incubator on Amp plates<br/>
 +
vector: psb1A3<br/>
 +
inserts: Not/ nOt/ noT + LOV-3A-PCR<br/>
==<span style="color:red;">red light receptor</span>==
==<span style="color:red;">red light receptor</span>==
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==<span style="color:orange;">Lysis cassette</span>==
==<span style="color:orange;">Lysis cassette</span>==
-
===NAME OF YOUR EXPERIMENT===
+
===New 2A assembly sequencing results===
-
'''Investigators:NAME'''
+
'''Investigators:Theo'''
 +
Results came in: [[File:S4+S15 no phosphatase.gb]]... The Part is finally there.. Stock was made
 +
As I found out during the next days the stock was not correct.. so the elusive part was again lost..
==<span style="color:grey;">Precipitator</span>==
==<span style="color:grey;">Precipitator</span>==

Latest revision as of 01:12, 22 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!


Contents

Meeting

attendants:
Time:

green light receptor

already done:


To-do:

blue light receptor

already done:


To-do:

red light receptor

already done:


To-do:

Lysis cassette

already done:


To-do:


Precipitator

already done:


To-do:

green light receptor

repeating quickchange PCR

Investigators:Julia
repeating PCR with fresh DNA template.
digest with DpnI,added directly to the PCR tube.
After NEB the DPNI has also a good activity in Phusion HF buffer.

blue light receptor

Ligation

Investigators: Sophie
Date: 25.08.11
continue from experiment: Digestion; Investigator: Sandra
Project Name: Blue light receptor
all samples stored in Blue light box


Transformation

Investigators: Sophie
Date: 26.08.11
Continue from Date: 26.08.11 Name: Sophie Experiment Ligation
Project Name: Blue light receptor

Procedure
1. take cells from -80°C freezer and put them on ice! (every eppi contains about 400 μl cells) 2. thaw cells on ice 20 minutes 3. pipette 50 μl cells and 2 μl DNA into eppi still on ice! 4. Incubate for 30 minutes on ice 5. Heat at 42°C for 60 sec 6. Incubate on ice for 5 minutes 7. Add 200 μl LB Broth 8. Incubate for 2 hours at 37°C (cells with lysis cassette at 30°C!!) 9. Plate 50 μl and 200μl on two different LB/Agar plates with appropriate antibiotic resistance


Documentation: Why are you doing this experiment? Name of the sample? Where are they stored? Name the vector with inserts, antibiotika resistance etc. Why? The last cloning was done with a NOT-PCR-Product which might make problems because it does not have enough bases after the restriciton sites.

Name of the samples: Not 1:1, Not 1:3, nOt 1:1 nOt 1:3, noT 1:1, noT 1: 3

stored in incubator on Amp plates

vector: psb1A3
inserts: Not/ nOt/ noT + LOV-3A-PCR

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

New 2A assembly sequencing results

Investigators:Theo


Results came in: File:S4+S15 no phosphatase.gb... The Part is finally there.. Stock was made As I found out during the next days the stock was not correct.. so the elusive part was again lost..

Precipitator

NAME OF YOUR EXPERIMENT

Investigators: NAME

                       

Retrieved from "http://2011.igem.org/Team:Freiburg/Notebook/26_August"