@prefix pplan: <http://purl.org/net/p-plan#> .
@prefix this: <http://purl.org/np/RAysi2SPEKm8TaYscHjRtWBr9tT0a2IA9d0jhRLRDP8bc> .
@prefix sub: <http://purl.org/np/RAysi2SPEKm8TaYscHjRtWBr9tT0a2IA9d0jhRLRDP8bc#> .
@prefix bs: <https://bioschemas.org/profiles/LabProtocol/0.4-DRAFT-2020_07_23/#> .
@prefix np: <http://www.nanopub.org/nschema#> .
@prefix pav: <http://purl.org/pav/> .
@prefix authority: <https://hypothes.is/a/> .
@prefix npx: <http://purl.org/nanopub/x/> .
@prefix xsd: <http://www.w3.org/2001/XMLSchema#> .
@prefix rdfs: <http://www.w3.org/2000/01/rdf-schema#> .
@prefix prov: <http://www.w3.org/ns/prov#> .
@prefix sp: <http://purl.org/net/SMARTprotocol#> .
@prefix dc: <http://purl.org/dc/elements/1.1/> .
sub:Head {
   this: np:hasAssertion sub:assertion ;
    np:hasProvenance sub:provenance ;
    np:hasPublicationInfo sub:pubInfo ;
    a np:Nanopublication .
}
sub:assertion {
   <http://purl.obolibrary.org/obo/CHEBI_17997> rdfs:label "liquid nitrogen" .
  <http://purl.obolibrary.org/obo/PO_0009005> rdfs:label "root" .
  <http://purl.obolibrary.org/obo/PO_0025034> rdfs:label "leaf" .
  sub:_1 rdfs:label "recalcitrant samples" .
  sub:_2 rdfs:label "recalcitrant samples" .
  sub:_3 rdfs:label "pestle" .
  sub:_4 rdfs:label "mortar" .
  sub:_5 rdfs:label "extraction buffer" .
  sub:step dc:description "Ground samples (leaf, shoot, root, and recalcitrant samples, approximately 0.5-1 g) using 1 ml of the extraction buffer with or without liquid nitrogen in mortar and pestle that are sterilized.CRITICAL STEP The procedure are carried out at room temperature except the centrifugation steps (at 4℃) as well as the time of precipitating of the nucleic acid using the isopropanol (on ice).CRITICAL STEP Severely disrupt the tissues to create the glaze mode of samples." ;
    sp:hasExperimentalInput <http://purl.obolibrary.org/obo/PO_0009005> , <http://purl.obolibrary.org/obo/PO_0025034> , sub:_1 , <http://www.ebi.ac.uk/efo/EFO_0000992> ;
    a pplan:Step ;
    bs:bioSampleUsed <http://purl.obolibrary.org/obo/PO_0009005> , <http://purl.obolibrary.org/obo/PO_0025034> , sub:_2 , <http://www.ebi.ac.uk/efo/EFO_0000992> ;
    bs:labEquipmentUsed sub:_3 , sub:_4 ;
    bs:reagentUsed <http://purl.obolibrary.org/obo/CHEBI_17997> , sub:_5 .
  <http://www.ebi.ac.uk/efo/EFO_0000992> rdfs:label "shoot" .
}
sub:provenance {
   sub:assertion pav:createdWith <https://nanotate.bitsfetch.com> ;
    prov:generatedAtTime "2021-03-05T11:34:30.681034"^^xsd:dateTime ;
    prov:wasAttributedTo <https://hypothes.is/users/iannotate> .
}
sub:pubInfo {
   sub:assertion prov:wasDerivedFrom authority:3HMP7H3GEeuJHqtY8IH5vg , authority:8vHAnH3GEeu_64_J_NTjhg , authority:AsPl1n3HEeuSaoMKfzNR1A , authority:U4k0VH3GEeuKIutEUEKDNA , authority:bwzvDn3GEeutc5OnvN9V8Q , authority:hB9uOn3GEeu1xVt7eD6q9A , authority:mxfzeH3GEeucLV_A-MvnNw , authority:rTnKzH3GEeuH5UcMOhvLdw , authority:winXQn3GEeudQt97vsqqag .
  sub:sig npx:hasAlgorithm "RSA" ;
    npx:hasPublicKey "MIGfMA0GCSqGSIb3DQEBAQUAA4GNADCBiQKBgQCEOtYac15zsWN/LAu3q7YFdW0UZNfX3BAarHW/LB1DHGcz0yYr0v7hKGbnCLJ+UkDfu3Mgj7BnCm1s8ulMULKfF4YjqS2YKHYZjNKVaIjSigFOpjOwIx99+qPlvd5CuG+Gia/sJA05jy2bPKLwb/kmVeeX0zORZudWLyRMLXp8UQIDAQAB" ;
    npx:hasSignature "eSiNGrBxHqXmxmmCqrlvmweZZTeW3VTUrw2lTNwq4K2WW1chx/PJ3CRcPlaXad5Wic4dQlwR/7H4c84jKVZo8IUwjwr2r6Xz48Ed9NE+3sVFVVzxf9PySoK8MkasyqBAHRrxOqL3wBaT9oNSczexTwaTkFveF3RvjMPANeaE7+Q=" ;
    npx:hasSignatureTarget this: .
  this: npx:introduces sub:step ;
    prov:generatedAtTime "2021-03-05T11:34:30.681034"^^xsd:dateTime ;
    prov:wasAttributedTo <https://hypothes.is/users/iannotate> .
}