TUDelft/12 September 2008

=September 12th=

Miniprep
We Miniprepped the parts that were colony PCRd and grown o/n (BBa_K115021, BBa_K115023 - BBa_K115026, BBa_K115028) yesterday. A Qiagen kit was used. All had a DNA concentration of about 150 ng/ul and a 260/280 of about 2.0.

Restriction
All Miniprepped parts were digested using XbaI and PstI, BB_R0080 was digested using EcoRI and SpeI, the empty vector pSB1AT3 was digested using XbaI and EcoRI. Restriction was performed for 3h @ 37°C with 10% of 10x buffer and 5 U of the restriction enzymes. Finally different amounts of DNA were used, as empty vector and R0080 will be used for all ligations, while the other parts will be used once. These restricted parts will be used for 3A assembly.

Ligation
The restricted parts will be ligated over the weekend at 16ºC.