{
  "@context": {
    "@vocab": "https://schema.org/"
  },
  "@type": "DigitalDocument",
  "@id": "https://raw.githubusercontent.com/BeBOP-OBON/TechOceanS_protocol_collection/main/odis_metadata/RPA_CRISPR-Cas_Assay.json",
  "hasDigitalDocumentPermission": "embargoed",
  "name": "CRISPR Cas – RPA Assay: Standard Operating Procedure",
  "description": "A standard operating procedure for the detection of eDNA from target species using a combined RPA/CRISPR-Cas protocol, which allows for the amplification of target-specific DNA, followed by its detection (fluorescence-based).",
  "url": "https://oceandecade.org/actions/better-biomolecular-ocean-practices/",
  "author": [
    {
      "@type": "Person",
      "name": "Fiona Bracken",
      "givenName": "Fiona",
      "familyName": "Bracken"
    },
    {
      "@type": "Person",
      "name": "Paola Drago",
      "givenName": " Paola",
      "familyName": "Drago"
    },
    {
      "@type": "Person",
      "name": "Anne Parle-McDermott",
      "givenName": " Anne",
      "familyName": "Parle-McDermott"
    },
    {
      "@type": "Person",
      "name": "Fiona Regan",
      "givenName": " Fiona",
      "familyName": "Regan"
    }
  ],
  "identifier": "",
  "keywords": [
    "CRISPR-Cas",
    "eDNA",
    "fluorescence-based detection",
    "amplicon",
    "Recombinase Polymerase Amplification",
    "SOP"
  ],
  "provider": {
    "@type": "Organization",
    "legalName": "Dublin City University",
    "name": "Dublin City University",
    "url": "https://www.dcu.ie/"
  },
  "license": "CC-BY",
  "publisher": {
    "@type": "Organization",
    "legalName": "Better Biomolecular Ocean Protocols",
    "url": " https://github.com/BeBOP-OBON"
  },
  "about": {
    "@type": "Action",
    "actionStatus": "PotentialActionStatus",
    "name": "Detection of eDNA from target species using a combined RPA/CRISPR-Cas protocol",
    "instrument": [
      "Thermo Block / Incubator",
      "Roche Light Cycler 480",
      "micropipettes",
      "centrifuge"
    ],
    "location": "This protocol can be applied in a laboratory equipped for molecular biology testing",
    "object": [
      "RPA kit",
      "Alt-R® A.s. Cas12a (Cpf1) V3, 500 µg",
      "Alt-R® A.s. Cas12a crRNA, 10 nmol (gRNA)",
      "Custom ssDNA-FQ Reporter",
      "Phosphate Buffered Saline",
      "Molecular grade water",
      "eDNA",
      "primers"
    ],
    "result": "Fluorescence emissions corresponding to DNA target sequences recognized by CRISPR-Cas12 mechanisms (FAM ƛex = 485 nm, ƛem = 535 nm)."
  }
}