... DMEM complete + 10 µg/mL polybrene by diluting 20 µL of 10 mg/mL polybrene into 20 mL media. Thaw the...with DMEM complete + 10 µg/mL polybrene, the final concentration of polybrene in each well should be 10 µg...glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Polybrene (10 mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 ...of the lentivirus in DMEM complete + 10 µg/mL polybrene. Note, this is just a sample of possible dilutions...Lentivirus (μL) Volume of DMEM complete + 10 µg/mL polybrene (µL) 0 0 500 1:5 300 200 1:10 150 350 1:50 30 ...solutions at various dilution. Make sure to use the polybrene-containing media to make the cell solution in ...total volume of 7 mL of DMEM complete + 10 µg/mL polybrene. Mix well by pipetting or inverting the tube. ...

...complete containing 10 μg/mL polybrene by diluting 20 μL of 10 mg/mL polybrene into 20 mL of media. If using...glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Polybrene (10 mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 ... before transduction. Note that in that case, polybrene should only be added at the time of viral transduction...lentivirus into DMEM complete containing 10 μg/mL polybrene: Dilution Volume of Lentivirus or Previous Dilution...) Volume of DMEM Complete Containing 10 μg/mL Polybrene (μL) Volume of Virus Added to Plate (μL) Volume... 9.45 mL of DMEM complete containing 10 μg/mL polybrene. Mix well by pipetting or inverting. Aliquot 1.35...cells may be more sensitive to lentivirus or to polybrene, in which case this incubation step can be reduced...

...μg/mL polybrene. TIP: Polybrene increases the efficiency of viral infection. However, polybrene is toxic.... Hexadimethrine Bromide (Polybrene) Prepare a 1mg/mL solution of polybrene (Sigma-Aldrich catalog #H9268... Vendor and catalog # Hexadimethrine bromide (polybrene)* Sigma-Aldrich: #H9268 Protamine Sulfate* MP ...your experiment Detailed protocols for preparing polybrene, protamine sulfate, and puromycin are located ... cell lines, substitute protamine sulfate for polybrene. c. Add lentiviral particle solution from step...at 4°C for up to one year. The powder form of polybrene is stable at 4°C for several years. Protamine ...

...EX0276-1 Benzonase 250 U/µl, Millipore #71205-3 Polybrene 10 mg/mL, Millipore, TR-1003-G Molecular Biology...300,000 cells/well in 1350 µL media and 11.1 µg/mL Polybrene into the wells containing the virus and the untransduced...,000 cells in 13.5 mL of media and 11.1 µg/mL Polybrene. Mix the cell suspension well before seeding. ...final volume in the well is 1.5 mL, and the final Polybrene concentration is 10 µg/mL. Mix well and incubate...

...glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Polybrene (10mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without...lentivirus into DMEM complete containing 10 μg/mL polybrene. Note, this protocol was developed using low titer...Volume of DMEM complete (μL) Volume of 10mg/mL polybrene (μL) 1:10 150 1348.5 1.5 1:25 60 1438.5 1.5 1:...

... this temperature but most antibiotics will not break down at this temperature. Check with your antibiotic...

...from the chamber. Use the metal spatula to gently break the gel cast open. Use a razor blade to cut the ...

... Solution in a well ventilated area and avoid breathing in the vapors. Workflow Timeline Day 1: Antigen...

...from the stable cell pool by trypsinization and break up any cell clumps into individual cells by passing...