Team:Freiburg/Notebook/4 July

From 2011.igem.org

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(Sequencing)
(3A-assembly: Digestion of plastic-binding site and GFP)
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step 2 to 4 was repeated 40x times.
step 2 to 4 was repeated 40x times.
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==<span style="color:grey;">3A-assembly: Digestion of plastic-binding site and GFP</span>==
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==<span style="color:grey;">Precipitator</span>==
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===3A-assembly: Digestion of plastic-binding site and GFP===
'''Investigators: Sophie, Sandra'''
'''Investigators: Sophie, Sandra'''

Revision as of 10:20, 15 July 2011

Contents

green light receptor

Digestion of PCR products

Investigators: Julia

Digest of PCR product Ccas.

Ccas:

  • Enzyme 1: XbaI
  • Enzyme 2: PstI

in vector pSB1C3:

  • Enzyme 1: SpeI
  • Enzyme 2: PstI

Comments: RFP is not digested with SpeI and PstI. It was digested again, that time with NEB4 and XbaI.

Lysis cassette

Sequencing

Investigators: Theo

PCR of Lys1, Lys2, Lys3, Lys4, Lys5.

Materials:

  • H2O
  • Seq. Buffer
  • DNA (Lys 1-Lys5)
  • Primer
  • Polymerase


PCR programm:

  1. 95°C 3 min
  2. 95°C 15 sec
  3. 50°C 15 sec
  4. 60°C 2 min
  5. 4°C stored til needed again

step 2 to 4 was repeated 40x times.

Precipitator

3A-assembly: Digestion of plastic-binding site and GFP

Investigators: Sophie, Sandra


  1. Digestion of GFP-plastic-binding site part (PCR product)
    • Enzyme 1: XbaI
    • Enzyme 2: SpeI
  1. Digestion of Promotor-RBS part (PR1-PR6)
    • EcoRI
    • SpeI
  1. Digestion of Vector pSB1K3
    • Enzyme 1: EcoRI
    • Enzyme 2: SpeI