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.