@prefix dce: <http://purl.org/dc/elements/1.1/> .
@prefix pplan: <http://purl.org/net/p-plan#> .
@prefix this: <http://purl.org/np/RAjeakz_EDD9glH9IAwz-_RBNN4e7FbZGOGv8kRGLt6Vk> .
@prefix sub: <http://purl.org/np/RAjeakz_EDD9glH9IAwz-_RBNN4e7FbZGOGv8kRGLt6Vk#> .
@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#> .
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 "cells" .
  sub:_2 rdfs:label "cells" .
  sub:_3 rdfs:label "haemocytometer" .
  sub:_4 rdfs:label "W.B.C. diluting fluid" .
  sub:_5 rdfs:label "10% FBS" .
  sub:_6 rdfs:label "1% of Pencillin-streptomycin solution" .
  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." ;
    sp:hasExperimentalOutput sub:_1 ;
    a pplan:Step ;
    bs:bioSampleUsed 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 "2021-03-05T10:05:28.013716"^^xsd:dateTime ;
    prov:wasAttributedTo <https://hypothes.is/users/iannotate> .
}
sub:pubInfo {
   sub:assertion prov:wasDerivedFrom authority:Azz5iH0pEeuRt5PWu1OTFQ , authority:Eg113H0pEeuSS69MJWdZSg , authority:HwcqEn0pEeuWMbM81jL9lQ , authority:Ndf9hH0pEeu1UsfKioflGA , authority:dGeaAH0pEeu9hrOzjMjrHQ , authority:kq-4WH0pEeucttstycv2Qg , authority:pxaVgn0pEeuyPfMwFWmvUQ .
  sub:sig npx:hasAlgorithm "RSA" ;
    npx:hasPublicKey "MIGfMA0GCSqGSIb3DQEBAQUAA4GNADCBiQKBgQCEOtYac15zsWN/LAu3q7YFdW0UZNfX3BAarHW/LB1DHGcz0yYr0v7hKGbnCLJ+UkDfu3Mgj7BnCm1s8ulMULKfF4YjqS2YKHYZjNKVaIjSigFOpjOwIx99+qPlvd5CuG+Gia/sJA05jy2bPKLwb/kmVeeX0zORZudWLyRMLXp8UQIDAQAB" ;
    npx:hasSignature "AT/JZAmX0G3TjJCp69mGsjbnEG4txmFhhuBdCEqmyA3punUx+34uJQlOFe8bZD5+6Qb7Gbg4iC5FBxVeNzkY0AL2R6vm70fshINABRWU1oLtSeRjyczmvwXXZpEnl6BnOqYhd/5t+T6aoYSRR1rZINQ6rK/Q/wzLJ6B0PQbHCsA=" ;
    npx:hasSignatureTarget this: .
  this: npx:introduces sub:step ;
    prov:generatedAtTime "2021-03-05T10:05:28.013716"^^xsd:dateTime ;
    prov:wasAttributedTo <https://hypothes.is/users/iannotate> .
}