Overexpression Stable Cell Line Generation

Case 1: Stable HLA-DQB2 overexpression HeLa Cell Line Generation

Backbone Maps:

Melt Curve:

Gene Expression:

Expression rate (%)

---

Case 2: The Human TREM2 and Human DAP12 genes were overexpressed in HEK293 Cell Line.

Plasmid information:

LV-EF1a>hTREM2/EGFP-PGK>Puro

Vector Map:

RT-qPCR:

Gene name	Primer name	Primer sequence		Fragment size
hGAPDH	hGAPDH-F		GGTGTGAACCATGAGAAGTATGA	123bp
	hGAPDH-R		GAGTCCTTCCACGATACCAAAG
hTREM2	hTREM2-F		CTGGAGATCTCTGGTTCCCC	218bp
	hTREM2-R		TTCACTGGGTGGATGTGTCC
hDAP12	hDAP12-F		GCGATTGCAGTTGCTCTACG	154bp
	hDAP12-R		TGTTTCCGGGTCGCTGC

Relative quantitative method was used to detect the expression change of the target gene, and the data was analyzed according to the 2-ΔΔCT method.

sample name	1	2	ΔCT (1-2)	ΔΔCT (experimental – control)	2-ΔΔCT	Expression efficiency
	hTREM2	hGAPDH
experimental	16.117	14.846	1.271	-18.41	348280.88	34828088%
control	34.758	15.077	19.681	0	1	100%

sample name	1	2	ΔCT (1-2)	ΔΔCT (experimental – control)	2-ΔΔCT	Expression efficiency
	hTREM2	hGAPDH
experimental	13.7	14.846	-1.147	-21.189	2390187.0	239018700%
control	35.119	15.077	20.042	0	1	100%

Dissolution curves of hTREM2, hDAP12 and hGAPDH: