Team:DTU-Denmark/Notebook/19 August 2013

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19 August 2013

Contents

lab 208


Main purpose


Who was in the lab


Kristian, Henrike, Julia

Procedure


Preparation of constructs to send for sequencing

Sec2, TAT3-2, TAT3-1a, CycAX and HAO constructs were prepared for sequencing.

Gradient PCR for creation of Biobricks

Made a screening to find good conditions for the extraction of our constructs for subsequent ligation into BioBricks.

PCR to linearize pZA21::RFP::araBAD

colony PCR on one of the Hello Wolrd constructs to confirm it's intact

Performed colony PCR on the construct containing Sec2 to check the template is correct because our PCRs on it are not working.

Results


gels

  • 1 kb ladder
  • N1 HF
  • N1 HF 2% DMSO 1 ml
  • N1 HF 5% DMSO
  • N1 HF 1 mB
  • N1 GC
  • N1 GC 2% DMSO
  • N1 GC 5% DMSO
  • N1 GC 5% DMSO 1 ml
  • N1 GC 5% DMSO 2 ml
  • N1 GC 1 mB
  • N2 HF
  • N2 HF 2% DMSO 1 ml
  • N2 HF 5% DMSO
  • N2 HF 1 mB
  • N2 GC
  • N2 GC 2% DMSO
  • N2 GC 5% DMSO
  • 1 kb ladder

(picture big gel)

  • 1 kb ladder
  • N2 GC 5% DMSO 1 ml
  • N2 GC 5% DMSO 2 ml
  • N2 GC 1 mB
  • pZA21::RFP::araBAD 0,5 uL MgCl2
  • pZA21::RFP::araBAD 1 uL MgCl2
  • pZA21::RFP::araBAD 2 uL MgCl2
  • negative pZA21::RFP::araBAD
  • col Sec2
  • col Sec2 (duplicate)
  • negative col Sec2
  • screening A1
  • screening A2
  • screening A3

(pic small gel)

Conclusion


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