Team:BIOTEC Dresden/Notebook2-1

1st September‘09:

1. Inoculation of 5 colonies from pTetFlp-KanR plasmid containing bacteria is sub-cultured into liquid agar and solid medium for overnight growth.

2. Minipreps of pRhaFlp-CmR clones has been done.

3. A Red/ET recombination has been done to insert PCR-CmR into pSC101-pRhaFlp plasmid and plated on Cm plates for overnight incubation.

2nd September‘09:

1. The concentrations of pRhaFlp-CmR miniprep DNAs has been done using nanodrop measuring device.

2. A restriction digest of parental pRhaFlp and pTetFlp plasmids along with pRhaFlp-CmR has been done with EcoRI and other minipreps of pRhaFlp-CmR with PstI.

3. Agarose Gel Electrophoresis is done to check the digests.

3rd September‘09:

1. A sequencing reaction has been set up for the PCR product of the plasmid pR6K-AmpSpec with Spec-forward and Spec-reverse primers using Taq polymerase.

2. Colonies from the pTetFlp-KanR are sub-cultured on new trimethoprin plates.

3. From Red/ET recombination of pRhaFlp clones positive colonies are reincubated from induced plate.

4th September‘09:

1. Minipreps of pRhaFlp-CmR clones has been done from the overnight cultures.

2. Re-electroporation of minipreps of 1, 12 and 13 of pRhaFlp-CmR has been done.