Lab Protocols Lab MaintenanceAutoclavingDishwashingChanging CO2 tanks on incubatorsCell CultureThawing mammalian cellsFreezing mammalian cellsAliquoting FBS, pen-strep, and L-glutamine (with FBS heat inactivation protocol)Cyro-Babies Label Template for AliquotingMammalian Cell Lysis & FractionationLysis buffer recipeLysing adherent mammalian cellsCell fractionation: isolation of nuclei Polyacrylamide Gel Electrophoresis & ImmunoblottingMicro-BCA protein concentration assay Mini gel castingMidi gel casting, electrophoresis, and blottingSDS-PAGE recipe NuPAGE western blot protocol Trans-Blot Turbo semi dry blottingWet tank transfer protocolImmunoprecipitation (IP)IP lysis buffer recipe Immunoprecipitation (IP) protocol Retroviral and Lentiviral Vectors for Cell EngineeringLentivirus preparationLentivirus transfection worksheet, 3rd generation packagingLentivirus transfection worksheet, 2nd generation packaging Lentivirus preparation (Fugene-assisted)Retrovirus preparation using Phoenix cells Retrovirus transfection worksheet Retrovirus preparation, VSVG pseudotyping Retrovirus preparation, (Fugene-assisted)Concentrating virus without ultracentrifugationTypical concentrations for eukaryotic antibiotic selectionsImmunofluorescenceCoverslip immunofluorescence Flow CytometryFlow Sorting PreparationCell Phenotypic AssaysGrowing cell spheroids in collagenSoft agar colony formation assayWound healing assay Ethanol fixation for cell cycle analysis Cell Culture Media RecipesMedia recipe for mouse PDAC cell lines Subculturing glioma stem cells (GSCs) Plasmid Manipulation, Propagation & AnalysisTransforming competent bacterial cell lines Transforming bacteria with pSLIK vectorsMaking LB agar plates Growing bacterial cultures Phenol-chloroform quick DNA extractionConcentrating DNA by precipitation in ethanol Reconstituting DNA from filter paper DNA gel electrophoresis Ordering Oligos (Thermo) & Geneblocks (Twist Biosciences)Plasmid SequencingPlasmidsaurus SequencingNEB Protocols for DNA WorkNEB® Primer Design ToolsPCR Using Q5® High-Fidelity DNA Polymerase (M0491)PCR Protocol for Phusion® High-Fidelity DNA Polymerase (M0530)NEBclonerDpn1 Digest ProtocolTime-Saver™ Qualified EnzymesHeat InactivationNEBioCalculatorNEBuilder HiFi DNA Assembly Reaction ProtocolTransformation ProtocolIsolation of RNA & qRT-PCRqRT-PCR using SYBR GreenQiagen RNeasy Mini Handbook Computational & Image Analysis ProtocolsHeatmap generation in MeV softwareR Studio code for PCA, Pearson, and heatmap workflow PLSR using SIMCA-PPLSR using RPLSR using MATLABPTM-SEA using RCell Profiler Tutorials PagePuncta quantification in ImageJProtocol for Spheroid AnalysisProtocol for Quantifying Protein Expression with Correlation PlotsProtocol for Puncta, Edge Intensity, Area, and Shape Analysis
