Molecular experiments notes of our project

Experiments overview spanning the project

June

6.21

1. Transformation of pMG36e into E.coli Top10

6.22

1. Transformation of pUC57-DRGN-1 into E.coli Top10
2. Grow pMG36e-transformed E.coli in LB broth

6.23

1. Isolation of pMG36e plasmid from bacteria solution
2. Grow DRGN-1 transformed E.coli in LB broth
3. Run agarose gel for pMG36e. Result: no target band

6.24

1. Isolation of pUC57-DRGN-1 from bacteria solution
2. Double-enzyme digestion of isolated pUC57-DRGN-1 using EcoRI and PstI.
3. Run agarose gel for confirmation. Result: confirmed.

6.25

1. Repeat the amplification experiment of pMG36e
2. Preparation of DH5ɑ competent cells

6.26

1. Select single colony of pMG36e-transformed E. coli, and transfer in LB broth.
2. Amplify the E. coli DH5ɑ containing pNZ8148 plasmid

6.27

1. The extraction of pMG36e and pNZ8148 plasmid
2. Digestion of pNZ8148 using XbaI
3. Agarose gel electrophoresis of digested pNZ8148. Result: confirmed, and the extracted plasmid is in super-coiled form

6.30

1. Gel electrophoresis for pMG36e, pUC57-DRGN-1 and pUC19. Results：no band for pMG36e, the other two are successful.

July

August

September

October

Interlab experiments

July

August

8.1
New chloramphenicol arrived. Tested competent cells again and compared new chloramphenicol with old one.
8.2
Negative results.
8.3
Tested chloramphenicol concentration from 1-50 μg/ml.
8.4
Negative results.
Tested chloramphenicol concentration from 50-200 μg/ml.
8.5
Negative results.
8.7
Tested borrowed chloramphenicol.
8.8
Positive results. Tested competent cells.
8.9
Positive results. Started Transformation.
8.10
Inoculated bacteria to liquid medium.
8.11
Diluted bacteria and measured OD₆₀₀ using spectrophotometer and measured fluorescence using plate reader.