NOTEBOOK
Gene synthesis at Kingsley Corporation
Order chloramphenicol, related antibodies, BCA kit
2017/6 / 20-2017 / 6/26Gene synthesis at Genescript
Start Interlab
Plasmid transformation, picking a single clone, amplification, determination of absorbance
2017/6 / 27-2017 / 7/3small amount of plasmid was extracted and sequenced
2017/7 / 4-2017 / 7/10Sequence is correct, the plasmid mentioned, get enough plasmid
2017/7 / 11-2017 / 7/17Resuscitation of Jurkat cells
2017/7 / 18-2017 / 7/24Jurkat cell proliferation, partial state is poor, discarded
Recovery another Jurkat cell
Recovery HEK293
2017/7 / 25-2017 / 7/31Continue to develop Jurkat and HEK293
2017/8 / 1-2017 / 8/7Lentivirus plasmid packaging in HEK293, virus particle extraction, concentration
The titer is low and does not meet the requirements
2017/8 / 8-2017 / 8/14Repeated packaging of the virus, transfection of Jurkat cells
Resistance screening
2017/8 / 15-2017 / 8/21RNA and protein extraction
Western blot with the flag antibody, the sample concentration is too large
2017/8 / 22-2017 / 8/28Protein Quantification BCA Kit Quantification of cell extracts and adjustment of sample volume.
Repeat Western blot
Reverse RNA to Cdna
2017/8 / 29-2017 / 9/4qRCR to verify the mRNA expression of foxp3, Usp7, SynNotch, Car
Learning and designing human blood flow sorting Treg cell technology
2017/9 / 5-2017 / 9-11Human blood extraction, Gradient centrifugation, Isolate monocytes
flow sorting, stimulating differentiation
2017/9 / 12-2017 / 9/18Treg cells were cultured and electrotransfection
2017/9 / 19-2017 / 9/25Flow analysis of transfected cells
Extraction of RNA
Protein extraction
2017/9 / 26-2017 / 10/2Reverse 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