Team:Freiburg/Notebook/15 July

From 2011.igem.org

(Difference between revisions)
(→NAME OF YOUR EXPERIMENT)
Line 59: Line 59:
<br/>
<br/>
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==<span style="color:green;">green light receptor</span>==
 
-
===NAME OF YOUR EXPERIMENT===
+
==<span style="color:orange;">Lysis cassette</span>==
-
'''Investigators:NAME'''
+
===Quick change===
-
==<span style="color:blue;">blue light receptor</span>==
+
'''Investigators: Theo'''
-
===NAME OF YOUR EXPERIMENT===
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==<span style="color:grey;">Precipitator</span>==
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'''Investigators:NAME'''
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'''PCR'''
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name:
 +
Ruediger
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Date:
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==<span style="color:red;">red light receptor</span>==
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18.07.2011
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===NAME OF YOUR EXPERIMENT===
+
|-
 +
| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Continue from Experiment (Date)PCR 1507
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'''Investigators:NAME'''
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(Name) Ruediger
 +
|-
 +
| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Project Name:
 +
GFP Pbd
 +
|}
 +
PCR-Mixture for one Reaction:
-
==<span style="color:orange;">Lysis cassette</span>==
+
For a 50 µl reaction use
-
===Quick change===
 
-
'''Investigators: Theo'''
+
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 32,5µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| H<sub>2</sub>0
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
-
==<span style="color:grey;">Precipitator</span>==
+
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 10µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 5x Phusion Buffer
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2.5µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Primer fw
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2.5µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Primer dw
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| dNTPs
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| DNA-Template
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 0.5 µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Phusion (add in the end)
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|}
 +
What program do you use?
 +
 
 +
10x (95C-41/52C-72C) + 25x ((95C-60C-72C)
 +
 
 +
 
 +
How did you label the PCR-Product, where is it stored and what do you do next?
 +
 
 +
 
 +
Reactions:
 +
 
 +
P28+P18+M14.1
 +
 
 +
P28+P19+M14.1
 +
 
 +
P28+P20+M14.1
 +
 
 +
 
 +
Results:
 +
 
 +
did not work well.
-
===NAME OF YOUR EXPERIMENT===
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Strange band size in lane 3. 4 and 5 did not work.
-
'''Investigators: NAME'''
+
[[Image:]][[Image:]]

Revision as of 14:54, 18 July 2011

Contents

Meeting

attendants: Tobi, Theo, Julia, Rüdiger, Jakob, Sandra

green light receptor

already done:


To-do:


blue light receptor

already done:

  1. Transformation of Lov-Tap in cells.


To-do:

  1. Cloning of promotor (medium) in Lov-Tap part and perform a transformation. Plating out of the cells on trypthophan-free medium.



red light receptor

already done:


To-do:

Lysis cassette

already done:

  1. repressor part (BBa_K098995) has no bases between RBS (B0034) and start codon (K098997), resulting in no translation.
    • quickchange of the repressor part to insert 6bp between RBS and start codon

To-do:

  1. DpnI digestion to digest the DNA template (methylated DNA) of the PCR to have only the new synthesized DNA strand.
  2. After digestion with DpnI, transformation of cells with the corrected part (BBa_K08995)


Precipitator

already done:


To-do:




Lysis cassette

Quick change

Investigators: Theo

Precipitator

PCR


Name:

Ruediger

Date:

18.07.2011

Continue from Experiment (Date)PCR 1507

(Name) Ruediger

Project Name:

GFP Pbd

PCR-Mixture for one Reaction:

For a 50 µl reaction use


32,5µl H20
10µl 5x Phusion Buffer
2.5µl Primer fw
2.5µl Primer dw
1µl dNTPs
1µl DNA-Template
0.5 µl Phusion (add in the end)

What program do you use?

10x (95C-41/52C-72C) + 25x ((95C-60C-72C)


How did you label the PCR-Product, where is it stored and what do you do next?


Reactions:

P28+P18+M14.1

P28+P19+M14.1

P28+P20+M14.1


Results:

did not work well.

Strange band size in lane 3. 4 and 5 did not work.

[[Image:]][[Image:]]

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