sub:assertion {
  <http://purl.obolibrary.org/obo/CHEBI_17992> rdfs:label "100mM sucrose" .
  <http://purl.obolibrary.org/obo/CHEBI_29234> rdfs:label "1% Triton -X" .
  <http://purl.obolibrary.org/obo/CHEBI_8984> rdfs:label "1% SDS" .
  sub:_1 rdfs:label "mosquitoes" .
  sub:_2 rdfs:label "mosquitoes" .
  sub:_3 rdfs:label "micropistle" .
  sub:_4 rdfs:label "1X STE buffer" .
  sub:_5 rdfs:label "20 µl/ ml  Proteinase K" .
  sub:_6 rdfs:label "10 µl/ ml  Rnase A" .
  sub:step dce:description "Grind mosquitoes  in  1.5 ml eppendorf  tube with micropistle in 50-100 µl 1X STE buffer ( 50mM Nacl,50mM Tris- HCL,100mM EDTA, Ph 8.0) along with 100mM sucrose. Add 1X STE buffer to a total volume of 300-500 µl for a single mosquito and 1 ml for mosquito pool like 4,6,8,10 numbers. Then add 1% SDS ,1% Triton -X , 10 µl/ ml  Rnase A (20mg/ml), 20 µl/ ml  Proteinase K (20mg/ml) and mix it." ;
    sp:hasExperimentalInput sub:_1 ;
    a pplan:Step ;
    bs:bioSampleUsed sub:_2 ;
    bs:labEquipmentUsed sub:_3 , <http://www.bioassayontology.org/bao#BAO_0010014> ;
    bs:reagentUsed <http://purl.obolibrary.org/obo/CHEBI_17992> , <http://purl.obolibrary.org/obo/CHEBI_29234> , <http://purl.obolibrary.org/obo/CHEBI_8984> , sub:_4 , sub:_5 , sub:_6 .
  <http://www.bioassayontology.org/bao#BAO_0010014> rdfs:label "1.5 ml eppendorf  tube" .
}