1. Since the results from our light experiments keep failing, some new ideas were computed. pSB1A2 is the backbone for our construct but since this is a high copy number plasmid perhaps the ratio of promotor to repressor is unequal (not enough repressor). A new enting in LC was done with ligX. These were grown for a few hours at 37°C and then put in 16°C room overnight so that more repressor can be made (its promotor is temperature-sensitive).
2. Ligation from 09/08 was electroporated in competent cells.
3. Miniprep of LigX, restriction digest and gel electrophoresis. The signals were good so a motherplate for ligX was made. Also, the vector pSB1A2 was extracted from the gel (39,1 ng/µl).

Nanodrop results:

• Gel electrophoresis of TER restriction
  • The vector for the TER is Psb1AK3 and has 3100bp, with an insert of ±3200 bp
  • Gel extraction

• Created a motherplate from EF1
• Ligated EF restriction + TER restriction
• Transferred the SAMS 1 and SAMS 2 colonies to liquid medium

• puc+A was plated
• X+K was again elctroporated