GRDZELISHVILI LAB PROTOCOLS (optimized and edited by Megan Moerdyk-Schauwecker)
LAB NOTEBOOK KEEPING:
GENERAL CELL CULTURE PROTOCOLS:
HPIV3 Titering Collection of Cells for Mycoplasma Testing Crystal Violet Staining Freezing cells HPDE and HPNE Mycoplasma Testing Passaging Cells Thawing PDA Cells Treating Cells for Mycoplasma
VIRUS TITRATION PROTOCOLS:
VSV Titering VSV Titering in 96 Well Format VSV Titering 24well VSV Titering in 12 Well Format_no overlay Vaccinia Titering Vaccinia Titering 96well plate SeV-wt Titering SeV-wt Titering in 96 Well Format Sev-wt Titering in 12-well format SeV-GFP Titering in 96 Well Format RSV Titering HSV1 Titering Adenovirus Titering in 96 Well Format HPIV3 Titering
VIRUS GROWTH AND PURIFICATION PROTOCOLS:
Growing rgRSV 102710 HSV-1 stock Infection of AD293 with Adenovirus for Virus Stocks Infection of BHK with VSV for Virus Stocks Infection of Vero with SeV-wt for Virus Stocks Infection of Vero with Vaccinia for Virus Stocks Purification of Adenovirus Purification of Vaccinia Purification of VSV_SeVIsolation and Purification of VSV RNPNegative Staining of virus for EMTreatment of VSV with ProK
PROTEIN ANALYSIS PROTOCOLS:
BRADFORD 96-well Coomassie Staining of gels Coomassie Staining of PVDF Ponceau Staining SDS PAGE gels TOTAL PROTEIN ISOLATION Western Blot 20cm Western Blot
RNA ANALYSIS PROTOCOLS:
Northern Blot 2 gels Northern Blot Synthesis of Radioprobe by 5′ End Labeling Synthesis of Radioprobe by In Vitro Transcription Trizol Isolation of RNASeV Genome TemplateIn vitro Synthesis of ssRNA_MarriottIn vitro transcriptionIsolation and Purification of VSV RNP
PCR PROTOCOLS:
VSV REVERSE GENETICS PROTOCOLS:
Plaque purification of VSV VSV Rescue Overview VSV Rescue using BSRT7 VSV Rescue using VVT7
MOLECULAR CLONING PROTOCOLS:
Competent Cells Gel extraction Transformation of Bacteria with Purified Plasmids Transformation of Plasmids into Bacteria
CELL VIABILITY AND APOPTOSIS PROTOCOLS:
Caspase3_AnnexinV MTT-LDH MTT Trypan blue TUNEL 2 chamber green label TUNEL 8-well chamber slide red label
STATISTICAL ANALYSIS PROTOCOL:
YEAST PROTOCOLS:
Extraction of Plasmids from Yeast and Transformation into Bacteria Preparing Competent Yeast Cells Preparing Yeast Samples for Western Blotting Transformation of Yeast Yeast glycerol stocks Yeast RNA Extraction by the Hot Phenol Method
OTHER PROTOCOLS:
Grdzelishvili Laboratory Protocols Coimmunoprecipitation (revision 1)Protein Isolation from Cultured Cells Sequence analysis of genomic RNA isolated from VSV virions Immunofluorescence Staining Protocol Immunofluorescence Protocol from Andrea Immunofluorescence staining procedure_3-23-10 Coomassie stainingCytoplasmic Extracts – Lysolecithin MethodHRP Immuno staining