sub:assertion {
sub:_1 rdfs:label "mosquitoes" .
sub:_10 rdfs:label "1% SDS" .
sub:_2 rdfs:label "1% Triton -X" .
sub:_3 rdfs:label "mosquitoes" .
sub:_4 rdfs:label "micropistle" .
sub:_5 rdfs:label "1.5 ml eppendorf tube" .
sub:_6 rdfs:label "1X STE buffer" .
sub:_7 rdfs:label "Proteinase K (20mg/ml)" .
sub:_8 rdfs:label "100mM sucrose" .
sub:_9 rdfs:label "Rnase A (20mg/ml)" .
sub:step dc: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 p-plan:Step ;
bs:bioSampleUsed sub:_3 ;
bs:labEquipmentUsed sub:_4 ,
sub:_5 ;
bs:reagentUsed sub:_10 ,
sub:_2 ,
sub:_6 ,
sub:_7 ,
sub:_8 ,
sub:_9 .
}