"
Team:HKUST/Notebook
From 2010.igem.org
14/06/2010
E.coli DH10B competent cells were successfully transformed with BioBrick
BBa_I746101 which contains agrC.
25/06/2010
PMH4 containing mCherry and PBI121 conatining gusA reporter gene were
successfully transferred into E.coli DH10B competent cells.
12/07/2010
Fusion PCR of agrC-mCherry was performed and products were confirmed by agarose
gel electrophoresis.
30/07/2010
Genomic DNA of Lactobacillus. Plantarum WCFS1 was extracted. Next, PCR of
plnB from the genomic DNA was conducted and products were confirmed by
agarose gel electrophoresis.
10/08/2010
Midi-prep of shuttle vector pMG36e was performed and extracted plasmids were
confirmed by enzyme digestion test.
16/08/2010
PCR of gusA from PBI121 was conducted and products were confirmed by agarose
gel electrophoresis.
17/08/2010
Fusion PCR of agrC-plnB was performed and products were confirmed by agarose gel
electrophoresis.
20/08/2010
Double-strand DNA of plnA promoter (124bp) was obtained. By following the
protocol of fusion PCR, annealing of the forward primer (79bp) with the reverse
primer (73bp) and the extension were accomplished.
26/08/2010
agrC-plnB was ligated into pBluescript KS (+). Ligation products were transferred
into E.coli DH10B competent cells by heat shock transformation. Ligation was
confirmed by colony PCR and plasmid digestion test.
13/09/2010
agrC was ligated into pBluescript KS (+). Ligation products were transferred
into E.coli DH10B competent cells by heat shock transformation. Ligation was
confirmed by colony PCR and plasmid digestion test.
24/09/2010
gusA was ligated into backbone pSB1C3. Ligation products were transferred
into E.coli DH10B competent cells by heat shock transformation. Ligation was
confirmed by colony PCR and plasmid digestion test.
