TUDelft/3 September 2008

=September 3rd =

3A Assembly
Today we started our first assembly attempt of a few bricks. We aim to produce our K115010 and K115011 using 3A assembly. We used the pSB1AC3 construction vector and cut the biobricks with the required sites (E&S or P&X). We performed an ethanol precipitation as found on OpenWetWare (listed on the protocols page) on the B0034 part as it is only 12 bp, the rest of the bricks was purified by a Qiagen Minelute. Concentrations after purification were all between 10-40 ng/ul, so we used 2.5 ul of the backbone and 5 ul of the inserts for a 15 ul ligation. The ratios were far from theoretical optimum, but we tried it anyway. Ligation was done o/n @ 16ºC.