Team:Peking/Notebook/TZZhu

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   Tianze Zhu's Notes
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My work is to find a way to achieve optimization of preservation of bioreporter bacteria. Utilizing preservation media and room temperature water pump to achieve a glassy form of bacteria sample and preserve in fridge.The other part of my work is to test and get the data of the dryed samples to see if the method has fulfilled our goal to preserve bacteria.

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Contents

Week 1st

Get the vector with Psal promoter and double digest it with X,P.

Get the insert of GFP gene and double digest it with S,P

Ligase the two part together in order to cnstruct the plasmid of Psal promoter and GFP,

Transfer the plasmid into trans 5a bacteria

Get the vector with Pbad promoter and double digest it with X,P.

Get the insert of GFP gene and double digest it with S,P

Ligase the two part together in order to cnstruct the plasmid of Pbad promoter and GFP,

Transfer the plasmid into trans 5a bacteria


Week 2nd

Get the vector with T7 promoter and double digest it with X,P.

Get the insert of GFP gene and double digest it with S,P

Ligase the two part together in order to cnstruct the plasmid of T7 promoter and GFP,

Transfer the plasmid into trans 5a bacteria


Week 3rd

Make the different types of protection media with gradient of trehalose PVP.


Week 4th

Use IPTG to induce the fresh cell containing plasmid of T7 promoter and GFP gene to express GFP.]

Detect it with flow cytometry assay and enzyme-linked immunoadsordent assay.

Determine the respone curve of the cell, and the intensity of GFP expression level.


Week 5th

Transfer the cells of T7 promoter and GFP, Mer promoter and GFP into protection media and dry them in the water pump. Store in fringe in 4°C.


Week 6th

ShangHai EXPO


Week 7th

Recover the stored cells and use cytometry assay and enzyme-linked immunoadsordent assay, the same methods used in fresh cells to determine the same parameter.


Week 8th

Recover the stored cells and use cytometry assay and enzyme-linked immunoadsordent assay, the same methods used in fresh cells to determine the same parameter.


Week 9th

Recover the stored cells and use cytometry assay and enzyme-linked immunoadsordent assay, the same methods used in fresh cells to determine the same parameter.


Week 10th

Collect the figure and data achieved in the previous three weeks to see how the condition and vigor of cells change.

Determine which kind of protection media is the best.