{"id":5185,"date":"2015-10-20T17:35:44","date_gmt":"2015-10-21T00:35:44","guid":{"rendered":"https:\/\/vermont.salk.edu\/?page_id=5185"},"modified":"2025-03-29T10:36:42","modified_gmt":"2025-03-29T17:36:42","slug":"instrumentation-services","status":"publish","type":"page","link":"https:\/\/www.salk.edu\/de\/science\/core-facilities\/viral-vector-core\/instrumentation-services\/","title":{"rendered":"Instrumentation &#038; Services"},"content":{"rendered":"<ul class=\"indent\">\n<li>Production of custom lentiviral, retroviral, AAV, G-deleted rabies, and adenoviral vectors<\/li>\n<li>\nProduction and distribution of stock lentiviral, retroviral, AAV, G-deleted rabies, and adenoviral vectors expressing reporter transgenes<\/li>\n<li>\nConsultation on experimental design, development and use of custom viral vectors<\/li>\n<li>\nProduction and distribution of AAV serotype kits useful for pilot studies<\/li>\n<\/ul>\n<h2>What to provide to the core:<\/h2>\n<ol style=\"padding-left:30px;\">\n<li>200ug (preferably >1ug\/ul) of your lentiviral or AAV plasmid. If large scale AAV production is requested, 500ug of AAV plasmid is required. DNA should have been purified using an endotoxin-free protocol (e.g. Endo-free maxi\/mega\/giga plasmid purification kits or equivalent). We do not accept miniprep purified DNA.<\/li>\n<li>Plasmid DNA should be checked for purity and have an A260\/280 of >1.8.<\/li>\n<li>AAV ITR plasmids should be checked for recombination by digestion with SmaI or XmaI. Each ITR contains two SmaI\/XmaI sites; digestion will cut out your insert. Excessive amounts of linearized full-length plasmid indicate recombination has occurred.<\/li>\n<li>Lentiviral and retroviral transfer plasmids should be confirmed by enzyme digest or sequencing.<\/li>\n<li>Please provide a gel image of your SmaI or XmaI digested AAV2 ITR plasmid, or your lentiviral transfer plasmid with the insert excised.<\/li>\n<li>To avoid recombination we recommend transforming and growing your lentiviral and AAV transfer plasmids in recombination deficient cells such as STBL3 @ 30\u00b0C IN 2XYT broth for no more that 16 hrs.<\/li>\n<li>Vector map and Sequence file, if available.<\/li>\n<li>Information about your gene of interest or insert any special requirements for handling: toxic, oncogenic, pro-apoptotic, etc.<\/li>\n<\/ol>\n<h2 style=\"margin-top: 40px;\">Recent Papers of Interest<\/h2>\n<ul class=\"indent\">\n<li><a href=\"http:\/\/www.pnas.org\/content\/107\/38\/16703.long\" target=\"_blank\" rel=\"noopener\">Selective viral vector transduction of ErbB4 expressing cortical interneurons in vivo with a viral receptor\u2013ligand bridge protein<\/a>., Choi et al., Proc Natl Acad Sci<\/li>\n<li><a href=\"http:\/\/www.cell.com\/neuron\/fulltext\/S0896-6273%2810%2900869-X\" target=\"_blank\" rel=\"noopener\">Monosynaptic Rabies Virus Reveals Premotor Network Organization and Synaptic Specificity of Cholinergic Partition Cells<\/a>. Stepiene et al., Neuron<\/li>\n<li><a href=\"http:\/\/www.ncbi.nlm.nih.gov\/pubmed\/21029739\" target=\"_blank\" rel=\"noopener\">The next step in gene delivery: Molecular engineering of adeno-associated virus serotypes.<\/a> Faust et al., J Mol Cell Cardio<\/li>\n<li><a href=\"http:\/\/www.ncbi.nlm.nih.gov\/pubmed\/19904235?itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum&amp;ordinalpos=3\" target=\"_blank\" rel=\"noopener\">Proteasome Inhibitors Decrease AAV2 Capsid derived Peptide Epitope Presentation on MHC Class I Following Transduction<\/a>. Finn et al., Mol Ther<\/li>\n<li><a href=\"http:\/\/www.ncbi.nlm.nih.gov\/pubmed\/19741632?itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum&amp;ordinalpos=1\" target=\"_blank\" rel=\"noopener\">The neonatal blood-brain barrier is functionally effective, and immaturity does not explain differential targeting of AAV9<\/a>. Saunders et al., Nat Biotech<\/li>\n<li><a href=\"http:\/\/www.ncbi.nlm.nih.gov\/pubmed\/19367261?itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum&amp;ordinalpos=8\" target=\"_blank\" rel=\"noopener\">Intravenous administration of self-complementary AAV9 enables transgene delivery to adult motor neurons.<\/a> Duque et al., Mol Ther<\/li>\n<li><a href=\"http:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC2835095\/?tool=pubmed\" target=\"_blank\" rel=\"noopener\">High-efficiency transduction of the mouse retina by tyrosine-mutant AAV serotype vectors.<\/a> Petrs-Silva et al., Mol Ther<\/li>\n<li><a href=\"http:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC2639530\/?tool=pubmed\" target=\"_blank\" rel=\"noopener\">Comparative analysis of HIV-1-based lentiviral vectors bearing lyssavirus glycoproteins for neuronal gene transfer<\/a>. Federici et al., Genet Vaccines Ther<\/li>\n<\/ul>\n<h2 style=\"margin-top: 40px;\">N\u00fctzliche Links<\/h2>\n<ul class=\"indent\">\n<li><a href=\"http:\/\/www.nature.com\/nprot\/journal\/v1\/n3\/abs\/nprot.2006.207.html\" target=\"_blank\" rel=\"noopener\">Production and characterization of recombinant adeno-associated viral vectors<\/a> &#8211; Samulski lab<\/li>\n<li><a href=\"http:\/\/www.addgene.org\" target=\"_blank\" rel=\"noopener\">Addgene<\/a> &#8211; distribution of useful plasmids including 2nd and 3rd generation lentiviral packaging systems.<\/li>\n<\/ul>","protected":false},"excerpt":{"rendered":"<ul","protected":false},"author":3,"featured_media":0,"parent":525,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"template-core-facilities-resources.php","meta":{"_acf_changed":false,"footnotes":""},"folder":[522],"class_list":["post-5185","page","type-page","status-publish","hentry"],"acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.3 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Instrumentation &amp; 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