慢病毒构建及功能试验

 STEP1 CLONING AND TRANSFORMATION（克隆和转化）
Estimated Time: 25-40 working days

Reagents and Materials	Number	Test/package	Experiment
DNA/RNA Extraction Kit	1	box	CLONING AND TRANSFORMATION
Plasmid Extraction Kit	1	box
Yeast Extract/Peptone	2	btl
Agar/Agarose	1	btl
T-vector	1	box
Restriction Enzme	4	pcs
DH5apha/BL21 E.Coli	1	str
Primer	4	pair
PCR/Taq Mix	2	pcs
Lenti-Virus Constructs***	1	set

*** provided by client
Results: 3 complete Lentiviral Construct (plasmid) containing GFP/BLANK/GFP+TARGET GENE*3
 STEP 2  TISSUE CULTURING AND TRANSFECTION（细胞组织培养和转染）
Estimated Time:15-20 working days

Reagents and Materials	Number	Test/package	Experiment
Hela/293T cell line	1	str	TISSUE CULTURING AND TRANSFECTION
RPMI 1640/DMEM	10	btl
Antibiotics (strep-kana)	5	mL
Tissue culture Plate	30	pcs
Fetal Bovine Serum	2	btl
Cytokines	1	Pcs
Transfection Reagent	1	pcs

Results: 3 complete Lentiviral transfected stable Hela cell lines containing GFP/BLANK/GFP+ TARGET Gene and Pseudovirus.
 STEP 3 FUNCTIONAL TEST (功能性实验)
5 complete gene/protein functional test by Western Blot, RT-PCR, Intracellular Fluorescence Microscopy, 2-Dimensional Proteomics, and Supernatant ELISA. （Westernblot，荧光定量PCR，胞内荧光染色，2-D电泳及蛋白组分析，免疫荧光或ELISA。）