Team:SUSTech Shenzhen/Materials

Materials

Double emulsion system: Opti-prep Density Gradient Medium, Isopropanol, Novec HFE 7500, Pico-Surf (TM) 1, 10ml, 2% in FC-40(Part number 3200202, Dolomite Corporation), TBST, PBS, DMEM with 10% Fetal Bovine Serum, 2% Sodium dodecyl sulfate(SDS).


Cell culture reagents: Recombinant Murine Wnt-3a(Peprotech Corporation), G418 Sulfate(Solarbio G8160), CHIR99021 drug, Union 293 Suspension medium.


Cell Counting: Fibro Media warm, Hemocytometer with cover slip, Eppendorf pipettes, 0.1% Trypan blue, Cell suspension, Microcentrifuge tube, Microscope.


Viral Infection: Regular growth media, 10mg/ml polybrene, (Un-) concentrated virus


Stable Cell Lines Construction: PEI, Opti-MEM, Drug-containing medium.


Western Blot: Wnt3a (C64F2) Rabbit mAb #2721(CST Corporation), 1x SDS Lysis buffer, RIPA Lysis buffer, Tris-buffered saline (TBS), 2x Laemmli sample buffer, Microcentrifuge tube, Plastic cell scrape, Molecular weight marker, 1x Transfer buffer, Cassette, Transfer tank, Ponceau S solution, Tris-Buffered Saline Tween (TBST), 3%BSA in TBST, Antibody, chemiluminescent substrate, CCD camera-based imager, Image analysis software.


Transfection: DNA expression vector, Plasmid DNA GeneXPlus Transfection Reagent, Serum-free growth medium, Antibiotics, Culture medium containing polyanions.


Mycoplasma Detection and treatment: Suspension or adherent cells, Sample Lysis Tube, Lysis buffer Agarose, TAE buffer, PCR product, Loading buffer, DNA marker, Ethidium bromide, Plasmocin™ Treatment(Invitrogen Corporation)


PCR: PCR Mix, PCR primers, ddH2O, Templates


Gibson Assembly: Fragments, Gibson Assembly Master Mix (2X), ddH2O, Thermocycler, NEB 5-alpha Competent E. coli cells.


Flow cytometry: ddH2O, Sodium Fluoride, Sodium Hypochlorite.