Fouad's used protocols and publications
General info on Schistosoma mansoni: https://www.angelfire.com/clone2/fouad/Schistosoma_mansoni.pdf
SmGPCR Datasheet (Full analysis of expressed il3 sequence) : https://www.angelfire.com/clone2/fouad/SmGPCR_datasheet.html
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pET System (Novagen)
pET vector manual (10th Ed.) https://www.angelfire.com/clone2/fouad/techniques/pET30a_manual.pdf
pET mechanism (summarized by Fouad): https://www.angelfire.com/clone2/fouad/techniques/pET_mechanism-Fouad.pdf
pET mechanism (Novagen): https://www.angelfire.com/clone2/fouad/techniques/pET_mechanism.pdf
pET30a-c map (Novagen): https://www.angelfire.com/clone2/fouad/techniques/pET30a-c_map.pdf
Sequencing data of pET30a and pET30a/il3-SmGPCR (cloned in DH5α): https://www.angelfire.com/clone2/fouad/techniques/Sequencing_of_pET30a__empty_plasmid__and_pET30a_il3-SmGPCR__DH5__.pdf
Alignment of pET30a (theoretical full sequence, Novagen) and sequenced one (experimental): https://www.angelfire.com/clone2/fouad/techniques/align_pET30alone_sequene_vs_full_pET30a__Novagen_.pdf
Alignment between pET30a-il3 (sequenced, after cloned in DH5α) and full sequence pET30a-il3 (software): https://www.angelfire.com/clone2/fouad/techniques/Alignment_between_pET30a-il3_sequenced__andl_full_sequence_pET30a-il3__.pdf
Rosetta/pET30a-il3 sequence (by Parasitology sequencer): https://www.angelfire.com/clone2/fouad/techniques/Rosetta_pET30a-il3_sequenced.html
Rosetta/pET30a-iL3 and DH5α-/pET30a-iL3 sequence: https://www.angelfire.com/clone2/fouad/techniques/Rosetta_pET30a-iL3.pdf
Alignment SmGPCR and S. japonicum GPCR congate: https://www.angelfire.com/clone2/fouad/techniques/Alignment_SmGPCR-SjGPCR__DNA___Protein_levels_.pdf
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FLAG-SmGPCR-EYFP (HEK E transfected cells):
https://www.angelfire.com/clone2/fouad/techniques/HEK293E_transfected_with_FLAG-SmGPCR-EYFP.pdf
Preparation of transfected HEK cell lysate and extract their GPCR: https://www.angelfire.com/clone2/fouad/techniques/Preparation_of_HEK_Cell_Lysate_and_extract_GPCR.pdf
Confocal Transfected HEK (FLAG-SmGPCR-GFP) procedure:
https://www.angelfire.com/clone2/fouad/techniques/confocal_HEK_procedure.pdf
RNAi on FLAG-SmGPCR-EYFP (HEK E transfected cells) using siPORT Amine and siPORT lipid transfection agents: https://www.angelfire.com/clone2/fouad/techniques/RNAi_on_SmGPCR_transfected_HEK.pdf
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α-Tubulin (S. mansoni, M80214): https://www.angelfire.com/clone2/fouad/techniques/_-Tubulin_.doc
Alignment between_tubulins (S. mansoni and human) https://www.angelfire.com/clone2/fouad/techniques/Alignment__between_tubulins.pdf
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Protocols
Conversion of µg of primer to picomoles: https://www.angelfire.com/clone2/fouad/techniques/Conversion_of__g_of_primer_to_pmoles_of_primer.pdf
6X PCR gel loading buffer: https://www.angelfire.com/clone2/fouad/techniques/6X_PCR_loading_buffer.pdf
Low mass DNA ladder: https://www.angelfire.com/clone2/fouad/techniques/low_mass_ladder.pdf
Amino acids table: https://www.angelfire.com/clone2/fouad/techniques/aminoacids.html
Protein determination RC & DC (microtitre BioRad): https://www.angelfire.com/clone2/fouad/techniques/Rc_Dc_microplate_assay.pdf
Nucleic acid precipitation by alcohols: https://www.angelfire.com/clone2/fouad/techniques/nucleic_acid_precipitation.mht
Miniprep protocol: https://www.angelfire.com/clone2/fouad/techniques/miniprep_protocol.html
Plasmid
Double Digestion of DNA (EcoR1 and Not1) https://www.angelfire.com/clone2/fouad/techniques/doubledigestionDNA.pdf
LB broth/medium preparation: https://www.angelfire.com/clone2/fouad/techniques/lbmedium_broth.html
Antibiotic for bacterial culture: https://www.angelfire.com/clone2/fouad/techniques/antibiotic_bacterial_culture.html
Preparation of competent cells: https://www.angelfire.com/clone2/fouad/techniques/preparation_of_competent_cells.html
Transformation in competent cells: https://www.angelfire.com/clone2/fouad/techniques/Transformation_of__competent_cells.pdf
Ligation using T4 ligase and Promega pGEM-T vector (white/blue colony screening): https://www.angelfire.com/clone2/fouad/techniques/promega_ligation.pdf
X-Gal/IPTG LB plates (white-blue screening): https://www.angelfire.com/clone2/fouad/techniques/X-Gal_LB_plate.pdf
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Silencer siRNA cocktail protocol: https://www.angelfire.com/clone2/fouad/techniques/silencercocktail.pdf
Silencer siRNA cocktail manual: https://www.angelfire.com/clone2/fouad/techniques/Silencer_cocktail_kit.pdf
Silencer siRNA labeling manual: https://www.angelfire.com/clone2/fouad/techniques/scilencer_labeling_kit.pdf
RNAi on schistosomula using siRNA: https://www.angelfire.com/clone2/fouad/techniques/Utilization_of_siRNA_on_schistosomula_SmGPCR.pdf
Two-step superscript RT-PCR: https://www.angelfire.com/clone2/fouad/techniques/2step_superscript_RTPCR.pdf
(A) Superscript III Reverse Transcriptase (RT): https://www.angelfire.com/clone2/fouad/techniques/superscriptIII_RT.pdf
(B) Superscript III Reverse Transcriptase (RT): https://www.angelfire.com/clone2/fouad/techniques/Reverse_transcriptase.doc
M-MLV RT (Invitrogen) protocol: https://www.angelfire.com/clone2/fouad/techniques/M-MLV.pdf
Sensiscript RT (Qiagen) manual: https://www.angelfire.com/clone2/fouad/techniques/sensiscript_RT__qiagen_.pdf
Real time RT-PCR + RNeasy Micro protocol: https://www.angelfire.com/clone2/fouad/techniques/Real_time_RT-PCR__RNeasy__protocol.pdf
Standard curve and melting curve in Real time PCR: https://www.angelfire.com/clone2/fouad/techniques/Real_time_PCR_standard_and_melting_curves_of_pCIneo-SmGPCR.pdf
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Treatment (Preclearing and preadsorbing) of anti SmGPCR IgG:
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Culture stuff
OPTI-MEM (reduced serum medium): https://www.angelfire.com/clone2/fouad/techniques/OPTI-MEM__reduced_serum_medium_.pdf
Culturing schistosomula in OPTi-MEM reduced serum: https://www.angelfire.com/clone2/fouad/techniques/OPTI-MEM_culturing_schistosomula.pdf
(How to prepare antibiotics and Schistosomula culture medium) https://www.angelfire.com/clone2/fouad/techniques/schistosomula_culture_antibiotics.pdf
Coating_slides_with_poly_L- lysine: https://www.angelfire.com/clone2/fouad/techniques/Coating_slides_with_poly_L-lysine.pdf
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Electrophoresis,Western Bolt, protein expression and IFA
Agarose gel electrophoresis: https://www.angelfire.com/clone2/fouad/techniques/Agarose_Gel_Electrophoresi1.pdf
Protein Expression in Prokaryotic system: https://www.angelfire.com/clone2/fouad/techniques/protein_expression_prokaryotes.pdf
His Bind Kit Manual: https://www.angelfire.com/clone2/fouad/techniques/His_Bind_Kit_manual.pdf
His Bind Kit (buffer preparation and regeneration): https://www.angelfire.com/clone2/fouad/techniques/His_Bind_Kit.pdf
SDS-PAGE: https://www.angelfire.com/clone2/fouad/techniques/SDS-PAGE.html
Elctrophoresis and Western Blot reagents: https://www.angelfire.com/clone2/fouad/techniques/electrophoresis_westernblot.html
Iodoacetamide and DTT stock preparation: https://www.angelfire.com/clone2/fouad/techniques/IAA_DTT_stocks.pdf
Immunoblot protocol from Chemicon: https://www.angelfire.com/clone2/fouad/techniques/Immunoblot_protocol.pdf
Iodoacetamide data sheet: https://www.angelfire.com/clone2/fouad/techniques/Iodoacetamide_Data_sheet.pdf
Western blot protocol: https://www.angelfire.com/clone2/fouad/techniques/Western_Immunoblotting_Protocol.html
Ponceau S solution (for reversible Western blot staining): https://www.angelfire.com/clone2/fouad/techniques/ponceau_S_solution.pdf
Stripping the PVDF protocol https://www.angelfire.com/clone2/fouad/techniques/stripping_Immunoblot.html
PVDF (BioRad manual): https://www.angelfire.com/clone2/fouad/techniques/PVDF_biorad.pdf
IEF microRotofor (BioRad) 0.1M H3PO4 preparation: https://www.angelfire.com/clone2/fouad/techniques/IEF_microRotofor__BioRad__10ml_of_0.1_H3PO4.pdf
ELISA protocol https://www.angelfire.com/clone2/fouad/techniques/elisaprotocol.html
ELISA plate template: https://www.angelfire.com/clone2/fouad/techniques/96_WELL_PLATE_DATA_SHEET.pdf
Tissue homogenization by RIPA buffer: https://www.angelfire.com/clone2/fouad/techniques/homogenization_in_RIPAbuffer.html
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Cleaning protein A column: https://www.angelfire.com/clone2/fouad/techniques/cleaning_proteinA.html
IgG purification by Protein A (affinity chromatography): https://www.angelfire.com/clone2/fouad/techniques/IgG_purification.html
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Immunofluorescent https://www.angelfire.com/clone2/fouad/techniques/Immunofluorescence_culturing_Schistosomula.pdf
Immunofluorescent using PFA fixative for Phalloidin/muscle identification: https://www.angelfire.com/clone2/fouad/techniques/Confocal_Immunofluorescence_using_PFA_as_a_fixative.pdf
Immunofluorescent using PFA fixative for Phalloidin/muscle identification (brief description): https://www.angelfire.com/clone2/fouad/techniques/brief_IFA.doc
Slide coating with poly L-lysine. https://www.angelfire.com/clone2/fouad/techniques/slide_coating_poly_lysine.pdf
Indirect Immunofluorescence (for in situ localization of SmGPCR in S. mansoni ): https://www.angelfire.com/clone2/fouad/techniques/insitulocalization.pdf
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RNA extraction and RNase inactivation:
DEPC treated stuff (for RNase inactivation): https://www.angelfire.com/clone2/fouad/techniques/DEPC.pdf
TriZol for RNA extraction: https://www.angelfire.com/clone2/fouad/techniques/Trizol.html
Trizol 2 for RNA extraction (from the current protocols of Molecular biology): https://www.angelfire.com/clone2/fouad/techniques/Trizol_RNA.html
RNeasy (Micro) Kit: https://www.angelfire.com/clone2/fouad/techniques/RNeasy.pdf
RNAlater RNA stabilization medium protocol: https://www.angelfire.com/clone2/fouad/techniques/RNAlater__Qiagen_.pdf
RNAlater (Qiagen protocol): https://www.angelfire.com/clone2/fouad/techniques/RNAlater__Qiagen_.pdf
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Crude adult S. mansoni antigen extraction:
RIPA buffer: https://www.angelfire.com/clone2/fouad/techniques/RIPA_buffer.html
Homogenization of the adult S. mansoni worms: https://www.angelfire.com/clone2/fouad/techniques/crude_Adult_S_mansoni_antigen.html
Experimentally infected mice with S. mansoni: https://www.angelfire.com/clone2/fouad/techniques/Experimentally_infected_mice_with_Schistosoma_mansoni.doc
Adult S. mansoni worm recovery from infected mice: https://www.angelfire.com/clone2/fouad/techniques/Mice_dissection_for_adult_S.htm
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Primer stuff
Real time master mix: https://www.angelfire.com/clone2/fouad/techniques/real_time_master_mix1.xls
Primer design: https://www.angelfire.com/clone2/fouad/techniques/primerdesign.pdf
(Full sequence of SmGPCR gene and EcoR1 and Not1 introduction) https://www.angelfire.com/clone2/fouad/techniques/SmGPCRfullgene.pdf
Primer stock preparation: https://www.angelfire.com/clone2/fouad/techniques/primer_stock_preparation.pdf
T7 promoter primer design: https://www.angelfire.com/clone2/fouad/techniques/newT7_promoter.doc
T7 promoter primers (RNAi) https://www.angelfire.com/clone2/fouad/techniques/T7promoter_primers.pdf
qRT-PCR primers for evaluating the RNAi of SmGPCR: https://www.angelfire.com/clone2/fouad/techniques/Primers_for_qRT.pdf
Strategy used to silence SmGPCR by RNAi: https://www.angelfire.com/clone2/fouad/techniques/strategy_of_RNAi_silencing_of_SmGPCR.pdf
Sense/Antisense qPCR SmGPCR #2: https://www.angelfire.com/clone2/fouad/techniques/quantitative_SmGPCR_primers__set__2_.pdf
Sense/Antisense qPCR Sm a-tubulin #3: https://www.angelfire.com/clone2/fouad/techniques/quantitative_Sm_tubulin_primers__set__3_.pdf
Sense/Antisense qPCR Hu B-actin #4: https://www.angelfire.com/clone2/fouad/techniques/quantitative_Human_beta_actin_primers__set__4_.pdf
Sense/Antisense qPCR Sm GAPDH: https://www.angelfire.com/clone2/fouad/techniques/S.mansoni_GAPDH_realtime_primers.pdf
RNAi of S. mansoni Dopamine 2 receptor 12404 (T7 primers and quantitative real-time primers): https://www.angelfire.com/clone2/fouad/techniques/SmDopa2__Sm12404__T7_primers_Results.pdf
RNAi of S. mansoni Metabotropic Glutamate receptor (T7 primers and quantitative real-time primers): https://www.angelfire.com/clone2/fouad/techniques/SmMGlu__10811__T7_primers_Results.pdf
RNAi of
Smp043340 &Smp043290: https://www.angelfire.com/clone2/fouad/techniques/RNAi_Smp043340_Smp043290.pdf
RNAi of Sm Tubulin-alpha (): https://www.angelfire.com/clone2/fouad/RNAi_Smp043340_Smp043290.pdf
Primers of Smp Histamine responsive gene like in S. mansoni: https://www.angelfire.com/clone2/fouad/techniques/primers_Smp_His_GPCR_related.pdf
Detection of integrity of dsRNA by gel electrophoresis: https://www.angelfire.com/clone2/fouad/techniques/Detection_of_the_integrity_of_dsRNA_by_gel_electrophoresis.pdf
Antibody and qPCR Delivery: https://www.angelfire.com/clone2/fouad/techniques/Antibody_Delivery.doc
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Technical Bulletins
RNA ppt by LiCl: https://www.angelfire.com/clone2/fouad/techniques/RNA_ppt_by_LiCl.pdf
Effect of freezing thawing of tissue on RNA integrity: https://www.angelfire.com/clone2/fouad/techniques/Effect_of_Freeze-Thawing_of_Tissue_on_RNA_Integrity.pdf
Avoiding DNA contamination of RT-PCR: https://www.angelfire.com/clone2/fouad/techniques/DNA_contamination_in_RT-PCR.pdf
The Basics of RNA isolation: https://www.angelfire.com/clone2/fouad/techniques/The_Basics_of_RNA_Isolation.pdf
Kozak sequence: https://www.angelfire.com/clone2/fouad/techniques/Kozak_sequence.doc
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Techniques
Hemocytometer (Cell counting chamber) and viability detection by Trypan blue: https://www.angelfire.com/clone2/fouad/techniques/Trypan_blue.pdf
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Antibody titer vs concentration: https://www.angelfire.com/clone2/fouad/techniques/Antibody_concentration_vs_titer.pdf
Centrifuge unit conversion: https://www.angelfire.com/clone2/fouad/techniques/centrifuge_unit_conversion_.xls
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Deglycosylation PNGase kit (Roche): https://www.angelfire.com/clone2/fouad/techniques/N-Glycosidase_F_Deglycosylation_Kit.pdf
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Acetone precipitation of proteins: https://www.angelfire.com/clone2/fouad/techniques/Acetone-precipitation.pdf
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Detegent facts: https://www.angelfire.com/clone2/fouad/techniques/Remove-detergent.pdf
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Phenol extraction of DNA: https://www.angelfire.com/clone2/fouad/techniques/phenol_extraction.doc
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TCA protein precipitation (1): https://www.angelfire.com/clone2/fouad/techniques/TCA_ppt_protocol.pdf
TCA protein precipitation (2): https://www.angelfire.com/clone2/fouad/techniques/TCA_protein_precipitation.doc
Trichloroacetic acid (protein ppt): https://www.angelfire.com/clone2/fouad/techniques/Trichloroacetic_acid_protein_ppt.pdf
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Gel_filtration_Sephadex_G-25_Medium: https://www.angelfire.com/clone2/fouad/techniques/Gel_filtration_Sephadex_G-25_Medium.pdf
Paraformaldehyde fixation: https://www.angelfire.com/clone2/fouad/techniques/Phalloidin-Rhodamine_Staining.pdf
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Seize_X_protein_G_manual: https://www.angelfire.com/clone2/fouad/techniques/Seize_X_protein_G_manual.pdf
IP of SmGPCR: https://www.angelfire.com/clone2/fouad/techniques/Seize_X_Mammalian_Immunoprecipitation_Kit.pdf
McGill Major Filled Form:
https://www.angelfire.com/clone2/fouad/Filled_Majors_only_-form-2005-2006.pdf
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Publication
(1st author) Prevalence of intestinal
helminthes of dogs and foxes from
(1st author) Diagnosis of canine echinococcosis: coproantigen versus necropsy in stray dogs and red foxes from
northern
(2nd author) Classical transmitters and their receptors in flatworms (2005): https://www.angelfire.com/clone2/fouad/techniques/review_GPCR.pdf
