Team:SSTi-SZGD/Notebook

March

3.11-3.17 Experiment part:

① Prepare DH5α competent cells
② Transform plasmid
③ Spreading and amplification

3.18-3.24 Experiment part:

① Extract plasmid
② Plasmid PCR validation
③ Learning Agarose gel electrophoresis

3.25-3.31 Experiment part:

① PAX01 plasmid resuscitation, amplification and validation

April

4.1-4.7 Experiment part:

① Preparation of B.S168 plasmid
② PAX01, PMA0911 and PDG1730 transformation (DH5α), amplification and validation

4.8-4.14 Experiment part:

① PAX01, PMA0911 and PDG1730 transformation (B.S168), amplification and validation
② PAX01, PMA0911 and PDG1730 plasmid extraction (DH5α)

4.15-4.21 Experiment part:

① Later practice of validation experiments, such as extraction and validation of protein by CTAB
② Practice PCR experiments

4.22-4.28 Experiment part:

① PAX01 electrotransformation (B.S168)
② PDG1730 validation (B.S168)

May

5.13-5.19 Experiment part:

① HA fermentation, segregation, purification, determination of concentration, determination of molecular weight and the validation of experimental protocol， etc.
② Hyaluronidase fermentation, segregation, extraction, enzyme activity detection and the validation of experimental protocol, etc.
③ PDG1730-HAase transformation, amplification and protein extraction

5.20-5.26 Experiment part:

① PDG1730-HAase’s SDS PAGE

June

6.10-6.16 Experiment part:

① PDG1730-HAase’s CBB (ultraviolet spectrophotometer)

6.17-6.23 Experiment part:

① PDG1730-HAase’s CBB (plate reader)
② PDG1730-HAase enzyme activity measurement (DNS method)

6.24-6.30 Experiment part:

① PDG173-HAase purification
② Biobrick: connect LHAase gene to PSB1C3, validation
③ interlab: Cell measurement protocol transformation, amplification and detection
④ interlab: OD600 Reference point

July

7.1-7.7 Experiment part:

① PDG1730-HAase purification
② Biobrick: connect LHAase gene to PSB1C3, validation
③ interlab: Cell measurement protocol transformation, amplification and detection OD600 Reference point
Particle Standard Curve
Fluorescence standard curve
CFU

7.8-7.14 Experiment part:

① PDG1730-HAase enzyme activity measurement (plate method)
② Puc57 plasmid extraction
③ interlab: Cell measurement protocol transformation, amplification and detection OD600 Reference point
Particle Standard Curve
Fluorescence standard curve
CFU

7.22-7.28 Experiment part:

① Biobrick: connect LHAase-sp gene to PSB1C3, validation

7.29-8.4 Experiment part:

① Biobrick: connect T1-hasA-RBS and phyla genes to PSB1C3

August

8.5-8.11 Experiment part:

① connect gtaB gene which is in the Puc57 plasmid to P43nmk noncarrier
② PMA0911 (contain target gene) transformation (B.S168), amplification and enzyme activity measurement by DNS method

8.12-8.18 Experiment part:

① High molecular weight HA segregation and purification
② use PMA0911, PDG1730 ELISA kit, nephelometry, DNS method to measure enzyme activity

8.19-8.25 Experiment part:

① PMA0911,PDG1730 (B.S168) plate method to measure enzyme activity
② High molecular weight HA molecular weight determination

September

9.2-9.7 Experiment part:

① PMA0911,PDG1730,gtaB,glmU transformation (B.S168E)
② HMW HA freeze-drying and delivery for test

9.8-9.15 Experiment part:

① PMA0911 (B.S168)purification
② GlmU, PMA0911 (BS168E) transformation , amplification, fermentationactivity

9.16-9.22 Experiment part:

① PDG1730 (BS168) transformation succeed, amplification, fermentation
② PDG1730 (B.S168) plate method to measure enzyme activity
③ PDG1730 (B.S168)purification

9.23-9.29 Experiment part:

① PMA0911(BS168) transformation , amplification, fermentation
② PMA0911 (B.S168) plate method to measure enzyme activity
③ PMA0911 (B.S168)purification

9.30-10.6 Experiment part:

① GlmU GlmS GtaB（B.S168E-PDG1730）transformation，amplification, fermentation
② GlmU GlmS GtaB (B.S168E-PDG1730)purification
③ LMW HA freeze-drying and delivery for test

Copyright © 2018 SSTi-SZGD All Rights Reserved.
Copyright © 2018 SSTi-SZGD All Rights Reserved.