Team:CPU CHINA/notebook

Home

Project

• Description
• Design
• Experiments
• Results
• Demonstrate

Human Practice

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

Parts

• Parts
• Basic Parts

Modeling

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

Lab

• Notebook
• Safety
• Interlab

Team

• Team
• Attributions
• Collaborations

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

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

Learning and designing human blood flow sorting Treg cell technology

2017/9 / 5-2017 / 9-11

Week13

Human blood extraction, Gradient centrifugation, Isolate monocytes

flow sorting, stimulating differentiation

2017/9 / 12-2017 / 9/18

Week14

Treg cells were cultured and electrotransfection

2017/9 / 19-2017 / 9/25

Week15

Flow analysis of transfected cells

Extraction of RNA

Protein extraction

2017/9 / 26-2017 / 10/2

Week16

Reverse the RNA into cDNA, quantify BCA protein, and verify the mRNA expression of Foxp3

Usp7, SynNotch and Car by qPCR, IP

2017/10 / 3-2017 / 10/10