...kinase promoter drives expression of puromycin. Puro R Puromycin resistance gene for selection of pLKO...sulfate, and puromycin are located in the “Appendix”. F.2. Determining the Optimal Puromycin Concentration...differently to puromycin selection. Addgene strongly recommends that you determine the optimal puromycin concentration...to fresh puromycin-containing media every other day. f. The minimum concentration of puromycin that results...with finer increments of puromycin to determine a more precise optimal puromycin concentration.) F.3. Protocol...expression of the puromycin resistance gene. f. To select for infected cells, add puromycin to the media at...positive control for the puromycin selection. Days 4+: g. Change to fresh puromycin-containing media as needed...

...°C. 0.1% Pluronic-F68 in PBS (A) Add 500 µL of 10% Pluronic-F68 to 49.5 mL PBS 0.01% Pluronic-F68 in PBS...Stocks of Pluronic F68 Pluronic F68, 10% solution A: 0.1% in PBS: 49.5 mL PBS + 500 µl Pluronic F68 B: 0.01%..., Hamilton 1X PBS pH 7.4 1X PBS-MK buffer 100X Pluronic-F68 NaCl MgCl 2 KCl Centrifugal filter units (...PBS (B) Add 5 mL of Buffer A to 45 mL PBS 0.001% Pluronic-F68 in PBS + 200 mM NaCl (C) (formulation buffer...Procedure Cover the filter membrane with 15 mL of 0.1% Pluronic F68 PBS and incubate for 10 min at RT Spin at ...and discard the flow through. Add 15 mL of 0.01% Pluronic F68 PBS and incubate for 10 min at RT. Spin at... discard the flow through. Add 15 mL of 0.001% Pluronic F68 + 200mM NaCl PBS. Spin at 3000 rpm for 5 min...

...Supermix for Probes no dUTP, Bio-Rad,1863023 10% Pluronic F-68, Thermo Fisher, 24040032 Droplet generation...Reagent Preparation 1X PCR Buffer containing 0.05% Pluronic F-68: Prepare immediately before use and vortex...using. 500 µL of GeneAmp 10X Buffer 25 µL of 10% Pluronic F-68 4475 µL Molecular Biology Grade Water Procedure...

... . The following day, cells were treated with puromycin. Colonies were allowed to grow in the presence...

... Linear, Transfection Grade, VWR 75800-188 10% Pluronic F-18, Thermo Fisher 24040032 Glutagro, Corning...BCD TFX 1000 mL BalanCD HEK293 Media 10 mL 10% Pluronic-F68 40 mL 200 mM Glutagro Do not add selective...

...deliver mammalian antibiotic resistance (e.g., puromycin, blasticidin), which enables selection of a stable...preparation Appropriate antibiotic for selection (e.g. puromycin, blasticidin) Reagent Preparation DMEM Complete...

...mM Sodium Citrate, pH 7.0 0.5% (wt/vol) N-laurosylsarcosine (Sarkosyl) 0.1 M 2-mercaptoethanol TRIzol®...