Team:Freiburg/Notebook/31 August

From 2011.igem.org

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{| style="border-spacing:0;"
{| style="border-spacing:0;"
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| See Digestion...
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| See Digestion...
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|}
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Transformation''' '''
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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: 31.08.11
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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: Ligation
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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: inducible promoter for pbd
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|}
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Procedure
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# take cells from -80°C freezer and put them on ice! (every eppi contains about 400 μl cells)
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# thaw cells on ice 20 minutes
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# pipette 50 μl cells and 2 μl DNA into eppi still on ice!
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# Incubate for 30 minutes on ice
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# Heat at 42°C for 60 sec
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# Incubate on ice for 5 minutes
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# Add 200 μl LB Broth
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# Incubate for 2 hours at 37°C (cells with lysis cassette at 30°C!!)
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# Plate 50 μl and 200μl on two different LB/Agar plates with appropriate antibiotic resistance
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'''Documentation:'''
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Why are you doing this experiment? Name of the sample? Where are they stored? Name the vector with inserts, antibiotika resistance etc.
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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: S54-ε 5-C3 1:1 / 1:3
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stored in incubator on Cm plates
|}
|}

Revision as of 15:35, 31 August 2011

Contents

green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

Miniprep

Investigators: Sandra

Miniprep of:

  • ♥-A3 Not 1
  • ♥-A3 noT 2
  • ♥-A3 noT 3

On the plates with tetracyclin nothing grew.


Testdigest

Investigators: Sandra

Testdigest of minipreps.

Digested with EcoRI and PstI.

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

Ligation


Name: Sophie Date: 31.01.11
Continue from Date: 30.08.11 Name: Sophie

Experiment: Digestion

Project Name: inducible promoter for pbd

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 S54 both
Y insert 2 ε 5
Z vector psb1C3
H2O

Documentation:

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


See Digestion...

Transformation


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

Experiment: Ligation

Project Name: inducible promoter for pbd

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.


Name: S54-ε 5-C3 1:1 / 1:3

stored in incubator on Cm plates

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