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Team:Freiburg/Notebook/20 July
From 2011.igem.org
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==<span style="color:grey;">Precipitator</span>== | ==<span style="color:grey;">Precipitator</span>== | ||
| - | + | '''Ligation''' | |
| - | ''' | + | |
| + | {| 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: 20.07 | ||
| + | |||
| + | |- | ||
| + | | colspan="2" style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Continue from Digestion Date 19.07 Name Ruediger | ||
| + | |||
| + | Experiment | ||
| + | |||
| + | |- | ||
| + | | 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 | ||
| + | |||
| + | |} | ||
| + | '''Procedure''' | ||
| + | |||
| + | |||
| + | 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 | ||
| + | | 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> | ||
| + | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Volume (μl) | ||
| + | |||
| + | |- | ||
| + | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| X insert 1 | ||
| + | | 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;"| -- | ||
| + | | 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;"| Y insert 2 | ||
| + | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| M14+P28+P18/19/20 | ||
| + | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2.7:1 | ||
| + | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2 each | ||
| + | |||
| + | |- | ||
| + | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Z vector | ||
| + | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| S39 / S40 | ||
| + | | 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 each | ||
| + | |||
| + | |- | ||
| + | | 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 | ||
| + | | 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;"| Ligation of 3 different GFP Pbd part (M14+P28+P18/19/20) into two vector with Promotor and RBS | ||
| + | |||
| + | |||
| + | S39 + M14+P28+P18 | ||
| + | |||
| + | S39 + M14+P28+P19 | ||
| + | |||
| + | S39 + M14+P28+P20 | ||
| + | |||
| + | |||
| + | S43 + M14+P28+P18 | ||
| + | |||
| + | S43 + M14+P28+P19 | ||
| + | |||
| + | S43 + M14+P28+P20 | ||
| + | |||
| + | |} | ||
Revision as of 15:42, 20 July 2011
Contents |
green light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
blue light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
Ligation
| Name: Ruediger | Date: 20.07 |
| Continue from Digestion Date 19.07 Name Ruediger
Experiment | |
| Project Name: GFP Pbd | |
Procedure
PCR tube:
total volume 20 μl
- add H2O (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
| Name of part | Ratio Insert:Vector
= 3:1 or 1:1 | Volume (μl) | |
| X insert 1 | -- | -- | -- |
| Y insert 2 | M14+P28+P18/19/20 | 2.7:1 | 2 each |
| Z vector | S39 / S40 | 2 each | |
| H2O |
Documentation:
Why are you doing this experiment? Where are your parts stored? Name the parts for ligation etc.
| Ligation of 3 different GFP Pbd part (M14+P28+P18/19/20) into two vector with Promotor and RBS
S39 + M14+P28+P19 S39 + M14+P28+P20
S43 + M14+P28+P19 S43 + M14+P28+P20 |
