@prefix dce: <http://purl.org/dc/elements/1.1/> .
@prefix pplan: <http://purl.org/net/p-plan#> .
@prefix this: <http://purl.org/np/RAtAKJJbfdpICi3gBVxevjeIaangOjOsWQNHEpSpZAULw> .
@prefix sub: <http://purl.org/np/RAtAKJJbfdpICi3gBVxevjeIaangOjOsWQNHEpSpZAULw#> .
@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#> .

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/OBI_0000918> rdfs:label "Dulbecco's modified eagle medium" .
  
  sub:_1 rdfs:label "cell suspension" .
  
  sub:_2 rdfs:label "cells" .
  
  sub:_3 rdfs:label "haemocytometer" .
  
  sub:_4 rdfs:label "W.B.C. diluting fluid" .
  
  sub:_5 rdfs:label "1% of Pencillin-streptomycin solution" .
  
  sub:_6 rdfs:label "10% FBS" .
  
  sub:step dce:description "Count cells by haemocytometer using W.B.C. diluting fluid: Add 10 µl of cell suspension to 190 µl of W.B.C. diluting fluid and mix well. Load the cell suspension in a haemocytometer and count the cells. Adjust cell concentration at 1 x 106 cells/ml with Dulbecco's modified eagle medium supplemented with 1% of Pencillin-streptomycin solution and 10% FBS. The approximate yield of cells from 4 ml of blood varies between 107-108.";
    a pplan:Step;
    bs:bioSampleUsed sub:_1, sub:_2;
    bs:labEquipmentUsed sub:_3;
    bs:reagentUsed <http://purl.obolibrary.org/obo/OBI_0000918>, sub:_4, sub:_5, sub:_6 .
}

sub:provenance {
  sub:assertion pav:createdWith <https://nanotate.bitsfetch.com>;
    prov:generatedAtTime "2021-02-14T05:26:39.967116"^^xsd:dateTime;
    prov:wasAttributedTo <https://hypothes.is/users/zuluaga.lizzz> .
}

sub:pubInfo {
  sub:assertion prov:wasDerivedFrom <https://hypothes.is/a/-jx25G6tEeujC5fYL3fuAw>,
      authority:4p0QXG6tEeuhbx_eqAb9Ng, authority:Eh2rSG6uEeuXkOPV2qhZUA, authority:NS5sYm6uEeubOlPUL_WTqQ,
      authority:g3l2Bm6tEeunnzMYDD96gQ, authority:laFEsm6tEeucxZcnEwg07A, authority:pMq1BG6tEeu075fQoAHsyw,
      authority:tzDryG6tEeua6H-ujqr03A .
  
  sub:sig npx:hasAlgorithm "RSA";
    npx:hasPublicKey "MIGfMA0GCSqGSIb3DQEBAQUAA4GNADCBiQKBgQCEOtYac15zsWN/LAu3q7YFdW0UZNfX3BAarHW/LB1DHGcz0yYr0v7hKGbnCLJ+UkDfu3Mgj7BnCm1s8ulMULKfF4YjqS2YKHYZjNKVaIjSigFOpjOwIx99+qPlvd5CuG+Gia/sJA05jy2bPKLwb/kmVeeX0zORZudWLyRMLXp8UQIDAQAB";
    npx:hasSignature "QkSWIrH3Rt+kyeLDja9Tw8vjXTTX0Akaeu4xr3LyaCa1fC19MnXVh0e88WeDwWnWHQFlKGgxQw11CemI5tXsJOOkkr8J/kspCXhgjUkfjTIc4vwQeVBGzvUN0h4+I95hVuGZDmh+MQGqZOrSlGzn7g7fIkWk2GRt1ZYd3hcSmns=";
    npx:hasSignatureTarget this: .
  
  this: npx:introduces sub:step;
    prov:generatedAtTime "2021-02-14T05:26:39.967116"^^xsd:dateTime;
    prov:wasAttributedTo <https://hypothes.is/users/zuluaga.lizzz> .
}