Difference between revisions of "Team:CPU CHINA/notebook"

 
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                 <div class="cd-timeline-content">
 
                 <div class="cd-timeline-content">
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                     <p>qRCR to verify the mRNA expression of foxp3, Usp7, SynNotch, Car</p>
+
                     <p>Continue to develop Jurkat</p>
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                     <p>Learning and designing human blood flow sorting Treg cell technology</p>
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                     <p></p>
 
                     <span class="cd-date">2017/9 / 5-2017 / 9-11</span>
 
                     <span class="cd-date">2017/9 / 5-2017 / 9-11</span>
 
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                 <div class="cd-timeline-content">
 
                 <div class="cd-timeline-content">
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                     <p>Human blood extraction, Gradient centrifugation, Isolate monocytes</p>
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                     <p>Continue to develop Jurkat and HEK293</p>
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                     <p>flow sorting, stimulating differentiation</p>
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                     <p>Transfect plasmids into Jurkat cells by Electrical transfection</p>
 +
                    <p>qRCR to verify the mRNA expression of foxp3, Usp7, SynNotch, CAR</p>
 
                     <span class="cd-date">2017/9 / 12-2017 / 9/18</span>
 
                     <span class="cd-date">2017/9 / 12-2017 / 9/18</span>
 
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                     <p>Treg cells were cultured and electrotransfection</p>
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                     <p>The cytokine stimulation experiment by IL-6 and IL17A</p>
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                    <p>Extraction of RNA</p>
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                    <p>Protein extraction</p>
 
                     <span class="cd-date">2017/9 / 19-2017 / 9/25</span>
 
                     <span class="cd-date">2017/9 / 19-2017 / 9/25</span>
 
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                     <p>Flow analysis of transfected cells</p>
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                     <p>Quantify BCA protein</p>
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                     <p>Extraction of RNA</p>
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                     <p>Western blot</p>
−
                    <p>Protein extraction</p>
+
 
                     <span class="cd-date">2017/9 / 26-2017 / 10/2</span>
 
                     <span class="cd-date">2017/9 / 26-2017 / 10/2</span>
 
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                 <div class="cd-timeline-content">
 
                 <div class="cd-timeline-content">
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                     <p>Reverse the RNA into cDNA, quantify BCA protein, and verify the mRNA expression of Foxp3</p>
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                     <p>Reverse the RNA into cDNA</p>
−
                     <p>Usp7, SynNotch and Car by qPCR, IP</p>
+
                     <p>Verify the mRNA expression of Foxp3，Usp7, SynNotch and CAR by qPCR, IP</p>
 
                     <span class="cd-date">2017/10 / 3-2017 / 10/10</span>
 
                     <span class="cd-date">2017/10 / 3-2017 / 10/10</span>
 
                 </div>
 
                 </div>

Latest revision as of 15:33, 30 October 2017

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