Template:Team:KULeuven/3 September 2009/VanillinProduction

NOT PCR

 * Electroporation of SAMS+TER ligation (from Tuesday ligation... the one that looked bad....)
 * Miniprepped the 4 colonies from SAMS
 * Restriction digest of SAMS
 * Gel electrophoresis of restriction from ech, fcs, sam5 and sam8
 * Result looks great (woohoo!)
 * Cut and purified from gel
 * Used ech and fcs to ligate using 5 &mu;l from ech and 23,5 &mu;l from fcs. There was not enough sam8 for ligation. Redo restriction at some point in the future...
 * 4 colonies from sam8 and ech were picked and plated

PCR

 * The PCR from yesterday went very well, we had a huge amount of amplified biobrick DNA of the different parts. Agarose gel electrophoresis was used to test if the different pieces of DNA created by PCR had the correct length.
 * Amplified DNA was purified and cut with different restriction enzymes. After restriction, DNA was purified again before ligation.
 * After the restriction, sam5, sam8 and terminator were ligated in a three-way ligation. The same was done for ech, fcs and terminator. We also started a ligation of just sam5 and sam8 and ech and fcs.