Team:Tuebingen/Notebook/Journal/Weekly

Week 26

Week 25

Week 24

Week 23

Week 22

Week 21

Week 20

Week 19

Week 18

Week 17

Week 16

Week 15

Week 14

Week 13

Week 12

Week 11

Week 10

Week 9

Week 8

Week 7

Week 6

Week 5

Week 4

Week 3

Week 2

We started out our iGEM project with several unsuccessful PCRs of Pfet3. We also tried restriction digests and ligations of Psuc2, rox1, mig1, and the mPRs into pSB1C3– without positive results. Unfortunately, we had massive problems (smiling marker) with staining our gelelectrophoresis gels thus had to start off our project with trouble-shooting. After some experiments we came to the conclusion that post-staining would be an expensive way to solve our gel problems. Due to better gels we could prove the successful PCRs of Panb1 and Psuc2 and promptly started ligation and transformation of these parts. Initial transformations of mig1 were unsuccessful. However, luc PCRs went very well and we already had highest hopes for this part. Unfortunately, we found an illegal restriction site inside of luc thus we had to come up with something special: site-directed-mutagenesis using 3 pairs of primers and Pfu-polymerase. Restricitons of Padh1, Psuc2, rox1, mig1, mPR Dr, mPR Xl and RFP (our initial reporter) went rather well but due to missing negative controls in the control gel further steps were not possible this week.