Date: 2023.07.21

Event: 1^st meeting at lab

Agenda: 

1.  LB medium preparation

2.  PCR amplification ( zliE-zliS, SacB)

3.  Electrophoresis verification: agarose gel

4.  Gel recycling

5.  Reverse PCR

 

Date: 2023.07.22

Event: 2^nd meeting at lab

Agenda:

1.  Continue to complete the remaining reverse PCR on the 21^st

2.  Complete the unsuccessful process on the 21^st PCR: primmer: zliE F1 + zliS R1

3.  Restriction digestion of empty plasmid and target fragment

4.  Enzyme digestion to verify EV ( empty vector )

5.  Recycle DNA

6.  Enzyme-linked plasmid fragments and target fragment

 

Date: 2023.07.23

Event: 3^rd meeting at lab

Agenda:

1.  PCR amplify RFP ( outline: prepare for RFP; plasmid 2--->plasmid 3 )

2. Homologous recombination ( homology arm )

3. Transform into CH5α E.coli and grow overnight.

 

Date: 2023.07.24

Event: 4^th meeting at lab

Agenda: 

1.  PCR ( E.coli with plasmid 3/plasmid 1 ( RFP without its own promoter ) )

2.  AMP solid medium

3.  Gel electrophoresis

4.  PCR ( to amplify the promoter of RFP ( PRFP ) )

5.  PCR ( Because the DNA electrophoresis of plasmid 1 failed in the previous test, plasmid 1 should be reconstructed )

 

Date: 2023.07.25

Event: 5^th meeting at lab

Agenda: 

1.  Gel electrophoresis

2.  Extract plasmid 3

3.  DNA extraction

4.  Digestion

5.  Enzyme link

6.  Convert

7.  Gel electrophoresis ( only for plasmid 1, since the previous ones failed )

8.  Culture E.coli with three different plasmids

 

Date: 2023.07.26

Event: 6^th meeting at lab

Agenda: 

1.  Colony PCR ( E.coli with plasmid 1/plasmid 2/plasmid 4 )

2.  AMP ( E.coli with plasmid 4; prepare for the next section: IPTG )

3.  Gel electrophoresis ( PCR product with plasmid 1&2&4 )

4.  Inverse PCR ( prepare for building up plasmid 3, amplifying plasmid 2 )

5.  Ampicillin configuration

 

Date: 2023.07.27

Event: 7^th meeting at lab

Agenda:

1.  IPTG & Sucrose control variable

2.  Gel electrophoresis

3.  RM media test

4.  Gel electrophoresis ( Because the IPCR of plasmid 2 failed in the previous section )

 

Date: 2023.07.28

Event: 8^th meeting at lab

Agenda:

1.  Extraction of sample fructan

2.  Testing the effect of sucrose and IPTG

3.  Prepare the solution for the standard curve of fructan

4.  Optical density value ( OD 540nm )

 

Date: 2023.0 9 . 01-09.08

Event:  Repeat some experiments under the guidance of mentor