The Notebook of

BioBrick Group

Finally we have to submit some biobricks that we made in our experiment. This notebook is focusing on constructing independent biobricks.There are about 30 biobricks that we plan to submit. In detail, we separate the into 4 groups:

Group one contains 8 different parts (Am4: Pcon+araC+pBAD; Am5: pRhl+RBS+mRFP+DT; Am6: pRHl/Las+RBS+GFP+DT; Am7: Pcon+RBS+LasR; Am8: RhlR+DT; Am9: RBS+amilCP+DT; Am10:RBS+amilCP+DT+Pcon+RBS; Am11: pRhl/Las+RBS);

Group two contains 8 different parts (Bm1: RBS+LasR+RhlR+DT; Bm2: Ptet+RBS+RhlR+DT; Bm3: Pcon+RBS_LasR+RhlR+DT; Bm4: pCin+CinR; Bm5: Pcon+RBS+Lac I; Bm6: pLac+RBS+luxI+DT; Bm7: RBS+CinI+DT; Bm8: pLux+RBS+LuxR+RBS+RhlI)

Week 1 8.21-8.24

1.We resuscitated the bacteria that contain Am4-Am11 parts.

2.We extracted all the plasmids and tested them by electrophoresis.

3.All the plasmids were cleaved by EcoR I and Pst I and we tested their qualities by electrophoresis.

4.We linked all the cleaved parts with pSB1C3 after we did the gel purification.

5.We transformed all the parts into E.coli. and extracted the plasmids which were tested by double cleavage.

6.We sent all the parts to sequence.

Week 2 8.25-8.31

1.We cleaved the Bm2, Bm6, and Bm7 plasmids to identify them.

2.We did the liquid culture for the bacteria containing Am4-Am11 plasmids.

3.We extracted the Am4-Am11 plasmids for storage.

4.We did the gel purification and linked the Bm2, Bm6, and Bm7 parts with pSB1C3.

5.We transformed our linkage products and extracted them.

6.We identified them by double cleavage and sent them to sequence.

Sichuan university