Notebook
Construction of Pasr and Pgada
4/26~4/27
- PCR Pasr and Pgada from E. coli MG1655 chromosome. Confirmed and extracted products by DNA gel.
4/28
- Digested the pSB1C3-sfGFP plasmid. Ligase the plasmid with Pasr / Pgada and RiboJ by PCR. Confirmed by DNA gel electrophoresis. Transformed the gene in DH5 alpha and spread the bacteria on LB plates.
4/30~5/1
- Confirmed the transformation by colony PCR ,enzyme digestion and DNA gel electrophoresis. Selected the correct colonies for sequencing and cultured them on LB plates.
5/9
- Store the correct colonies in glycerol in -80O C.
6/1
- Transformed each plasmid into BL21(DE3)
Construction of Pasr and Pgada
5/9
- Prepared the M9 medium in different pH value.
5/11
- Pre-cultured the DH5 alpha-pSB1C3 -Pasr-sfGFP strain.
5/12
- Cultured the DH5 alpha-pSB1C3 -Pasr-sfGFP strain in M9 medium in different pH value. Measured the OD600 value and fluorescence every hour.
5/18
- Pre-cultured DH5 alpha-pSB1C3 –Pgada-RiboJ-sfGFP strain.
- Prepared the M9 medium in different pH value.
5/19
- Cultured the DH5 alpha-pSB1C3 –Pgada-RiboJ-sfGFP strain in M9 medium in different pH value. Measured the OD600 value and fluorescence every hour.
6/2~6/3
- Repeated the experiment on 5/11~5/12
7/29
- Pre-culture the BL21(DE3)-pSB1C3 –Pasrr-sfGFP strain
7/30
- Cultured the BL21(DE3)-pSB1C3 –Pasr -sfGFP strain on 96 well plate in different pH value. Measure the fluorescence every 5 minutes in 30 minutes.
9/4
- Pre-culture the BL21(DE3)-pSB1C3 –Pgada-RiboJ-sfGFP strain
9/5
- Cultured the BL21(DE3)-pSB1C3 –Pgada-RiboJ-sfGFP strain strain on 96 well plate in different pH value. Measure the fluorescence every 5 minutes in 30 minutes.