Team:UC Chile/Cyano/Labbook/week21

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Cyanolux & SpiderColi - Pontificia Universidad Católica de Chile, iGEM 2012



Monday:

  • Band purification of LuxCD & Lux EG.
  • Gel electrophoresis: Colony PCR of RS2 cut (no extra restriction site) and Luc CDEG. Right sized bands: 11 and 12. Plasmids from preceding colonies were extracted and sized was checked by electrophoresis. Finally they were ligated to B0014 and B0015.

Tuesday:

  • PCR Run

VB psb1C3, VBpsb1A3, VB psb1T3, VB psb1K3, V psb1K5, Lux CD

  • Colony PCR

RS1+RS2: (12 colonies selected) B0014+RS2: (12 colonies selected) B0015+RS2: (12 colonies selected) Lux CD+Lux EG: (4 colonies selected) Positive and negative control

Results: correct controls. Most of the colonies were right sized. Error in Lux CD+EG because the bands were much smaller than expected (1000 bp). Two colonies were chosen in each case. Digestions left overnight: Lux CD from Brick, and Lux EG obtained by PCR. Wrong digestions were eliminated.

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