@prefix obo: <http://purl.obolibrary.org/obo/> .
@prefix dce: <http://purl.org/dc/elements/1.1/> .
@prefix pplan: <http://purl.org/net/p-plan#> .
@prefix this: <http://purl.org/np/RAHi9L2Ftk2pQ_lSeWbu23B7opN3IajkEAm8CAs35q7eE> .
@prefix sub: <http://purl.org/np/RAHi9L2Ftk2pQ_lSeWbu23B7opN3IajkEAm8CAs35q7eE#> .
@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 {
  obo:OBI_0400155 rdfs:label "water bath" .
  
  sub:_1 rdfs:label "100 mg/ml lysozyme solution" .
  
  sub:step dce:description """Bring down temperature of suspension to ~30 °C and add 20 µl of 100 mg/ml lysozyme solution (final concentration, 2 mg/ml) to it. Incubate suspension at 37 °C for 6 h to overnight in a water bath.
Note: In case of fast-growing mycobacteria such as Mycobacterium smegmatis (M. smegmatis), go directly to step 7. However, if DNA is to be isolated from slow-growing species, follow steps 4-6 which involve lysis of cells by a physical method. Because of enhanced disruption of cells, this would significantly enhance yield.""";
    a pplan:Step;
    bs:bioSampleUsed <http://www.bioassayontology.org/bao#BAO_0002104>;
    bs:labEquipmentUsed obo:OBI_0400155;
    bs:reagentUsed sub:_1 .
  
  <http://www.bioassayontology.org/bao#BAO_0002104> rdfs:label "suspension" .
}

sub:provenance {
  sub:assertion pav:createdWith <https://nanotate.bitsfetch.com>;
    prov:generatedAtTime "2021-03-20T05:39:47.164703"^^xsd:dateTime;
    prov:wasAttributedTo <https://hypothes.is/users/iannotate> .
}

sub:pubInfo {
  sub:assertion prov:wasDerivedFrom authority:G3LVuojzEeu_bQeA7ItEzQ, authority:MVp9QojzEeuIwvdaiT_htQ,
      authority:WyamZIjzEeuUay-2mYqjfg, authority:cyo8HIjzEeuG1Gc-5ZGJtg .
  
  sub:sig npx:hasAlgorithm "RSA";
    npx:hasPublicKey "MIGfMA0GCSqGSIb3DQEBAQUAA4GNADCBiQKBgQCEOtYac15zsWN/LAu3q7YFdW0UZNfX3BAarHW/LB1DHGcz0yYr0v7hKGbnCLJ+UkDfu3Mgj7BnCm1s8ulMULKfF4YjqS2YKHYZjNKVaIjSigFOpjOwIx99+qPlvd5CuG+Gia/sJA05jy2bPKLwb/kmVeeX0zORZudWLyRMLXp8UQIDAQAB";
    npx:hasSignature "b8dX4Z3HdbViHuM7OrEfmaDCOtdttLtr0ZLCbapt3HsT2wCBkRZ65uSCufE0VO/yZt5W9VvyI/26TGvRxlN7uBve1+r+YhPi17vQ947pXt/+FAxFBzxNoKC4mbxyKjm/0hii0UxzGH62G1dt/hwSzKzG5maO9o+GasJWm+v4psI=";
    npx:hasSignatureTarget this: .
  
  this: npx:introduces sub:step;
    prov:generatedAtTime "2021-03-20T05:39:47.164703"^^xsd:dateTime;
    prov:wasAttributedTo <https://hypothes.is/users/iannotate> .
}