Team:CPU CHINA/Notebook

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Project

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• Design
• Experiments
• Results
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Human Practice

• Silver Hp
• Intergrated and Gold Hp
• Education and Public Engagement

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• Basic Parts

Modeling

• Overview
• Interactions between cytokines Model
• Reciprocal Differentiation Model
• Exploration Model

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NOTEBOOK

Week1

Gene synthesis at Kingsley Corporation

Order chloramphenicol, related antibodies, BCA kit

2017/6 / 20-2017 / 6/26

Week2

Gene synthesis at Genescript

Start Interlab

Plasmid transformation, picking a single clone, amplification, determination of absorbance

2017/6 / 27-2017 / 7/3

Week3

small amount of plasmid was extracted and sequenced

2017/7 / 4-2017 / 7/10

Week4

Sequence is correct, the plasmid mentioned, get enough plasmid

2017/7 / 11-2017 / 7/17

Week5

Resuscitation of Jurkat cells

2017/7 / 18-2017 / 7/24

Week6

Jurkat cell proliferation, partial state is poor, discarded

Recovery another Jurkat cell

Recovery HEK293

2017/7 / 25-2017 / 7/31

Week7

Continue to develop Jurkat and HEK293

2017/8 / 1-2017 / 8/7

Week8

Lentivirus plasmid packaging in HEK293, virus particle extraction, concentration

The titer is low and does not meet the requirements

2017/8 / 8-2017 / 8/14

Week9

Repeated packaging of the virus, transfection of Jurkat cells

Resistance screening

2017/8 / 15-2017 / 8/21

Week10

RNA and protein extraction

Western blot with the flag antibody, the sample concentration is too large

2017/8 / 22-2017 / 8/28

Week11

Protein Quantification BCA Kit Quantification of cell extracts and adjustment of sample volume.

Repeat Western blot

Reverse RNA to Cdna

2017/8 / 29-2017 / 9/4

Week12

Continue to develop Jurkat

2017/9 / 5-2017 / 9-11

Week13

Continue to develop Jurkat and HEK293

Transfect plasmids into Jurkat cells by Electrical transfection

qRCR to verify the mRNA expression of foxp3, Usp7, SynNotch, CAR

2017/9 / 12-2017 / 9/18

Week14

The cytokine stimulation experiment by IL-6 and IL17A

Extraction of RNA

Protein extraction

2017/9 / 19-2017 / 9/25

Week15

Quantify BCA protein

Western blot

2017/9 / 26-2017 / 10/2

Week16

Reverse the RNA into cDNA

Verify the mRNA expression of Foxp3，Usp7, SynNotch and CAR by qPCR, IP

2017/10 / 3-2017 / 10/10