Team:Freiburg/Notebook/4 July

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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
  • 100ng DNA (Lys 1-Lys5)
  • 2ng Primer
  • Polymerase
  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.