Team:Freiburg/Notebook/4 July

From 2011.igem.org

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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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'''Investigators: Sophie, Sandra'''

Revision as of 10:01, 14 July 2011

Contents

23. Labday


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.


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.


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

Investigators: Sophie, Sandra