Team:Freiburg/Notebook/1 September

From 2011.igem.org

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==<span style="color:grey;">Precipitator</span>==
==<span style="color:grey;">Precipitator</span>==
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===NAME OF YOUR EXPERIMENT===
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===Ligation===
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 +
'''Ligation'''
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{| style="border-spacing:0;"
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name: Sophie
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Date: 1.9.11
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 +
|-
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| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Continue from Date: 31.08.11 Name: Sophie
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 +
Experiment: Digestion
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 +
|-
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| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Project Name:
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|}
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'''Procedure'''
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 +
 
 +
PCR tube:
 +
 
 +
total volume 20 μl
 +
 
 +
 
 +
# add H<sub>2</sub>O (17 μl -X-Y-Z)
 +
# add 2 μl Ligase Buffer 10x
 +
# add Insert 1, Insert 2(when proceeding from 3A digestion use 2 μl of each)
 +
# add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)
 +
# Add 1 μl T4-DNA Ligase
 +
# Incubate 10-30 min at room temperature
 +
# heat for 20 minutes at 80°C
 +
# store at -20°C or directly proceed to transformation
 +
 
 +
 
 +
{| style="border-spacing:0;"
 +
| 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;"| Name of part
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Ratio Insert:Vector
 +
 
 +
<nowiki>= 3:1 or 1:1</nowiki>
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Volume (μl)
 +
 
 +
|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| X insert 1
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| GFP-pbd-PCR a/b
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| both
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Y vector
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| pGEX
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| H<sub>2</sub>O
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| style="background-color:#bfbfbf;border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="background-color:#bfbfbf;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;"|
 +
 
 +
|}
 +
'''Documentation:'''
 +
 
 +
Why are you doing this experiment? Where are your parts stored? Name the parts for ligation etc.
 +
 
 +
 
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Why? To get data about the plastic binding for modeling
 +
 
 +
stored in: “Minipreps, verdaut”-box
 +
 
 +
ligation-products stored in: ligation box
 +
 
 +
parts for ligation: GFP-pbd-PCR a/b, pGEX
 +
 
 +
|}
 +
 
-
'''Investigators: NAME'''
 
{{:Team:Freiburg/Templates/footer}}
{{:Team:Freiburg/Templates/footer}}

Revision as of 12:15, 1 September 2011


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

Contents

green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

Picking clones

Investigators: Sandra

There were several clones on the cm plates. Clones were picked and incubated in LB cm medium at 37°C.

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

Ligation

Ligation


Name: Sophie Date: 1.9.11
Continue from Date: 31.08.11 Name: Sophie

Experiment: Digestion

Project Name:

Procedure


PCR tube:

total volume 20 μl


  1. add H2O (17 μl -X-Y-Z)
  2. add 2 μl Ligase Buffer 10x
  3. add Insert 1, Insert 2(when proceeding from 3A digestion use 2 μl of each)
  4. add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)
  5. Add 1 μl T4-DNA Ligase
  6. Incubate 10-30 min at room temperature
  7. heat for 20 minutes at 80°C
  8. store at -20°C or directly proceed to transformation


Name of part Ratio Insert:Vector

= 3:1 or 1:1

Volume (μl)
X insert 1 GFP-pbd-PCR a/b both
Y vector pGEX
H2O

Documentation:

Why are you doing this experiment? Where are your parts stored? Name the parts for ligation etc.


Why? To get data about the plastic binding for modeling

stored in: “Minipreps, verdaut”-box

ligation-products stored in: ligation box

parts for ligation: GFP-pbd-PCR a/b, pGEX


                       

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