{"id":5425,"date":"2015-10-25T20:45:30","date_gmt":"2015-10-26T03:45:30","guid":{"rendered":"https:\/\/vermont.salk.edu\/?page_id=5425"},"modified":"2025-02-05T13:18:50","modified_gmt":"2025-02-05T21:18:50","slug":"resources","status":"publish","type":"page","link":"https:\/\/www.salk.edu\/es\/science\/core-facilities\/resources\/","title":{"rendered":"Instrumentaci\u00f3n y servicios"},"content":{"rendered":"

Equipment<\/h2>\r\n

Nikon Eclipse TE 300 and Diaphot 300 Inverted Microscopes
\r\nNikon SMZ1000 Dissection Microscopes
\r\nOlympus CKX31 and SZX7 microscopes
\r\nNarishige MO-188 and MN-188 hydraulic micromanipulators
\r\nHamilton Thorne XYCLone Laser System
\r\nEppendorf PiezoXpert Microinjector
\r\nNanoDrop 2000 Spectrophotometer
\r\nVibration-free Tables
\r\nPLI-90 Microinjectors
\r\nSutter Instrument P-97 Pipette Puller
\r\nAlcatel Microforge
\r\nMyCycler System
\r\nBio-Rad Gene Pulser Electroporator
\r\nVAPRO Vapor Pressure Osmometer
\r\nBench shaker
\r\nForma 3110 CO2 Incubators with copper interior
\r\nNuair 425-400 HEPA filter vertical laminar flow hoods
\r\nBio Cool III Controlled-rate Freezer
\r\n-80\u00b0C freezer
\r\n-20\u00b0C freezer
\r\nMVE 600 liquid nitrogen cryo-preservation storage systems
\r\nUVP Biodoc-It Image system<\/p>\r\n\r\n

Services<\/h2>\r\n\r\nCRISPR\/Cas9 microinjections<\/strong>\r\n\r\nDetails\u00bb<\/a>\r\n
\r\n
\r\n\r\nSince 2013, Transgenic Core has been successfully using CRISPR\/Cas9 nuclease technology for efficient and precise generation of gene edited mouse models. This technology is a powerful tool that allows production of knock-out, knock-in and point mutant mouse lines in only 3\u20134 months. Transgenic Core will provide consultation, design editing strategy and assist with genotyping.\r\n
<\/div>\r\n\r\n
<\/div>\r\nSubmit CRISPR microinjection request<\/a>\r\n
<\/div>\r\nOrder CRISPR reagents:
\r\n \r\n1.Cas9 Protein
\r\nIDT
\r\nAlt-R\u00ae S.p.HiFi Cas9 Nuclease V3
\r\n100 ug
\r\nCat. No 1081060
\r\nhttps:\/\/www.idtdna.com\/pages\/products\/crispr-genome-editing\/alt-r-crispr-enzymes<\/a>\r\n
<\/div>\r\n2. sgRNAs (1.5 mol of each)
\r\nSynthego
\r\nhttps:\/\/www.synthego.com\/success\/synthetic-sgrna<\/a>\r\n
<\/div>\r\n3. ssDNA (3 ug)
\r\n
<\/div>\r\na) IDT
\r\nhttps:\/\/www.idtdna.com\/pages\/products\/genes-and-gene-fragments\/megamers-single-stranded-dna-fragments<\/a>\r\n
<\/div>\r\nb) Genewiz:
\r\nhttps:\/\/www.genewiz.com\/en\/Public\/Services\/Gene-Synthesis\/Single-Stranded-DNA-Synthesis<\/a>\r\n
<\/div>\r\nc) Genescript:
\r\nhttps:\/\/www.genscript.com\/new-single-stranded-dna-synthesis-service.html?page_no=1&position_no=1&sensors=googlesearch<\/a>\r\n
<\/div>\r\n\r\n\r\n<\/div>\r\n<\/div>\r\n\r\n
\r\n\r\nDNA microinjection<\/strong>\r\n\r\nDetails\u00bb<\/a>\r\n
\r\n
\r\n\r\nTransgenic models have proved to be a powerful experimental tool in mammalian genetics. DNA microinjection is the most common method to produce random transgene integration into the mouse genome.\r\n
<\/div>\r\nThe Transgenic Core performs all the steps necessary to produce transgenic mice, including DNA purification, superovulation of donor females, mating set-up, embryo harvesting, microinjection and embryo transfer to recipient females.\r\n
<\/div>\r\nThe investigator is responsible for supplying the DNA construct with all vector sequences removed. Prokaryotic cloning vector sequences inhibit the expression of eukaryotic genes introduced into the mouse genome. We microinject DNA into the male pronucleus of a minimum of 200 mouse embryos in order to provide the investigator with at least 20 pups. Generally, a round of microinjections produces 2-6 founders. Pups will be weaned, tailed and numbered by Core personnel.The investigator will need to perform DNA isolation and analyze the DNA by PCR and Southern blotting.Transgenic founders will be transferred to the investigator holding room at 4 to 5 weeks of age.\r\n
<\/div>\r\nTo request DNA microinjection service:<\/strong><\/em>\r\n
<\/div>\r\n