{"id":8,"date":"2012-03-21T17:12:50","date_gmt":"2012-03-21T17:12:50","guid":{"rendered":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/?page_id=8"},"modified":"2016-06-13T00:52:35","modified_gmt":"2016-06-13T00:52:35","slug":"news","status":"publish","type":"page","link":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/news\/","title":{"rendered":"Lab Protocols"},"content":{"rendered":"

GRDZELISHVILI LAB PROTOCOLS (optimized and edited by\u00a0Megan Moerdyk-Schauwecker)<\/em><\/strong><\/p>\n

 <\/p>\n

LAB NOTEBOOK KEEPING:<\/p>\n

Lab Notebook Keeping<\/a><\/p>\n

 <\/p>\n

GENERAL CELL CULTURE PROTOCOLS:<\/p>\n

HPIV3 Titering<\/a> Collection of Cells for Mycoplasma Testing<\/a> Crystal Violet Staining<\/a> Freezing cells<\/a> HPDE and HPNE<\/a> Mycoplasma Testing<\/a> Passaging Cells<\/a> Thawing PDA Cells<\/a> Treating Cells for Mycoplasma<\/a><\/p>\n

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VIRUS TITRATION\u00a0PROTOCOLS:<\/p>\n

VSV Titering<\/a> VSV Titering in 96 Well Format<\/a> VSV Titering 24well<\/a> VSV Titering in 12 Well Format_no overlay<\/a> Vaccinia Titering<\/a> Vaccinia Titering 96well plate<\/a> SeV-wt Titering<\/a> SeV-wt Titering in 96 Well Format<\/a> Sev-wt Titering in 12-well format<\/a> SeV-GFP Titering in 96 Well Format<\/a> RSV Titering<\/a> HSV1 Titering<\/a> Adenovirus Titering in 96 Well Format<\/a> HPIV3 Titering<\/a><\/p>\n

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VIRUS GROWTH AND PURIFICATION\u00a0PROTOCOLS:<\/p>\n

Growing rgRSV 102710<\/a> HSV-1 stock<\/a> Infection of AD293 with Adenovirus for Virus Stocks<\/a> Infection of BHK with VSV for Virus Stocks<\/a> Infection of Vero with SeV-wt for Virus Stocks<\/a> Infection of Vero with Vaccinia for Virus Stocks<\/a> Purification of Adenovirus<\/a> Purification of Vaccinia<\/a> Purification of VSV_SeV<\/a>Isolation and Purification of VSV RNP<\/a>Negative Staining of virus for EM<\/a>Treatment of VSV with ProK<\/a><\/p>\n

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PROTEIN ANALYSIS\u00a0PROTOCOLS:<\/p>\n

BRADFORD 96-well<\/a> Coomassie Staining of gels<\/a> Coomassie Staining of PVDF<\/a> Ponceau Staining<\/a> SDS PAGE gels<\/a> TOTAL PROTEIN ISOLATION<\/a> Western Blot 20cm<\/a> Western Blot<\/a><\/p>\n

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RNA ANALYSIS\u00a0PROTOCOLS:<\/p>\n

Northern Blot 2 gels<\/a> Northern Blot<\/a> Synthesis of Radioprobe by 5′ End Labeling<\/a> Synthesis of Radioprobe by In Vitro Transcription<\/a> Trizol Isolation of RNA<\/a>SeV Genome Template<\/a>In vitro Synthesis of ssRNA_Marriott<\/a>In vitro transcription<\/a>Isolation and Purification of VSV RNP<\/a><\/p>\n

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PCR PROTOCOLS:<\/p>\n

RT-PCR<\/a> PCR<\/a><\/p>\n

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VSV REVERSE GENETICS\u00a0PROTOCOLS:<\/p>\n

Plaque purification of VSV<\/a> VSV Rescue Overview<\/a> VSV Rescue using BSRT7<\/a> VSV Rescue using VVT7<\/a><\/p>\n

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MOLECULAR CLONING\u00a0PROTOCOLS:<\/p>\n

Competent Cells<\/a> Gel extraction<\/a> Transformation of Bacteria with Purified Plasmids<\/a> Transformation of Plasmids into Bacteria<\/a><\/p>\n

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CELL VIABILITY AND APOPTOSIS\u00a0PROTOCOLS:<\/p>\n

Caspase3_AnnexinV<\/a> MTT-LDH<\/a> MTT<\/a> Trypan blue<\/a> TUNEL 2 chamber green label<\/a> TUNEL 8-well chamber slide red label<\/a><\/p>\n

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STATISTICAL ANALYSIS PROTOCOL:<\/p>\n

Statistical Analysis<\/a><\/p>\n

 <\/p>\n

YEAST PROTOCOLS:<\/p>\n

Extraction of Plasmids from Yeast and Transformation into Bacteria<\/a> Preparing Competent Yeast Cells<\/a> Preparing Yeast Samples for Western Blotting<\/a> Transformation of Yeast<\/a> Yeast glycerol stocks<\/a> Yeast RNA Extraction by the Hot Phenol Method<\/a><\/p>\n

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OTHER PROTOCOLS:<\/p>\n

Grdzelishvili Laboratory Protocols<\/a>\u00a0Coimmunoprecipitation (revision 1)<\/a>Protein Isolation from Cultured Cells<\/a> Sequence analysis of genomic RNA isolated from VSV virions<\/a> Immunofluorescence Staining Protocol<\/a> Immunofluorescence Protocol from Andrea<\/a> Immunofluorescence staining procedure_3-23-10<\/a> Coomassie staining<\/a>Cytoplasmic Extracts – Lysolecithin Method<\/a>HRP Immuno staining<\/a><\/p>\n

 <\/p>\n

 <\/p>\n","protected":false},"excerpt":{"rendered":"

GRDZELISHVILI LAB PROTOCOLS (optimized and edited by\u00a0Megan Moerdyk-Schauwecker)   LAB NOTEBOOK KEEPING: 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\u00a0PROTOCOLS: VSV Titering VSV Titering in […]<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"open","ping_status":"open","template":"","meta":{"jetpack_post_was_ever_published":false,"footnotes":""},"class_list":["post-8","page","type-page","status-publish","hentry"],"jetpack_shortlink":"https:\/\/wp.me\/P2j6yj-8","jetpack_sharing_enabled":true,"_links":{"self":[{"href":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/wp-json\/wp\/v2\/pages\/8","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/wp-json\/wp\/v2\/comments?post=8"}],"version-history":[{"count":14,"href":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/wp-json\/wp\/v2\/pages\/8\/revisions"}],"predecessor-version":[{"id":416,"href":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/wp-json\/wp\/v2\/pages\/8\/revisions\/416"}],"wp:attachment":[{"href":"http:\/\/pages.charlotte.edu\/valery-grdzelishvili\/wp-json\/wp\/v2\/media?parent=8"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}