Microscopy:Article Title: In vivo Mitophagy Monitoring in Caenorhabditis elegans to Determine Mitochondrial Homeostasis
Article Snippet: .. Greiner Petri dishes (60 x 15 mm) (Greiner Bio One International, catalog number: 628161) Microscope slides 75 x 25 x 1 mm (Marienfeld-Superior, catalog number: 10 006 12) Microscope cover glass 18 x 18 mm (Marienfeld-Superior, catalog number: 01 010 30) Use the following transgenic nematodes to monitor mitophagy: IR1631: N2; Ex003 [p myo-3 TOMM-20::Rosella; pRF4], R1284: N2; Is [p myo-3 mtGFP]; Ex011 [p lgg-1 DsRed::LGG-1; p myo-2 GFP]) Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center) 70% of EtOH Potassium dihydrogen phosphate (KH 2 PO 4 ) (EMD Millipore, catalog number: 104873) di-Potassium hydrogen phosphate (K 2 HPO 4 ) (EMD Millipore, catalog number: 137010) Sodium chloride (NaCl) (EMD Millipore, catalog number: 106404) Peptone (BD, BactoTM, catalog number: 211677) Streptomycin (Sigma-Aldrich, catalog number: S6501) Agar (Sigma-Aldrich, catalog number: 05040) Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01) Calcium chloride dehydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: C5080) Magnesium sulfate (MgSO 4 ) (Sigma-Aldrich, catalog number: M7506) Nystatin stock solution (Sigma-Aldrich, catalog number: N3503) di-Sodium hydrogen phosphate (Na 2 HPO 4 ) (EMD Millipore, catalog number: 106586) Levamisole (Sigma-Aldrich, catalog number: L9756) Paraquat (Sigma-Aldrich, catalog number: 856177) Carbonyl cyanide m-chlorophenylhydrazone (CCCP) (Sigma-Aldrich, catalog number: C2759) Dimethyl sulfoxide cell culture grade BC (DMSO) (AppliChem, catalog number: A3672,0250) Phosphate buffer (1 M; sterile, see Recipes ) Nematode growth medium (NGM) agar plates (see Recipes ) M9 buffer (see Recipes ) Levamisole (0.5 M, see Recipes ) M9-levamisole solution (20 mM solution, see Recipes ) Paraquat (0.5 M, see Recipes ) Carbonyl cyanide m-chlorophenylhydrazone (49 mM; CCCP, see Recipes ) .. UV crosslinker (Vilber Lourmat, model: BIO-LINK – BLX-E365) Zeiss AxioImager Z2 epifluorescence microscope (Zeiss, model: Zeiss AxioImager Z2) Olympus DP71 CCD camera (Olympus, model: Olympus DP71) Zeiss AxioObserver Z1 confocal microscope (Zeiss, model: Zeiss AxioObserver Z1) Dissecting stereomicroscope (Olympus, model: SMZ645) Incubators for stable temperature (AQUA ® LYTIC incubator 20 °C)
Article Title: Protein Synthesis Rate Assessment by Fluorescence Recovery after Photobleaching (FRAP)
Article Snippet: .. Greiner Petri dishes (60 × 15 mm) (Greiner Bio One, catalog number: 628161) 35 mm plates (Corning, catalog number: 430165) Microscope slides 75 × 25 × 1 mm (Marienfeld-Superior, catalog number: 10 006 12) Microscope cover glass 18 × 18 mm (Marienfeld-Superior, catalog number: 01 010 30) C. elegans strains (wild type [N2], ife-2[ok306] , N2; Ex [p ife-2 GFP, pRF4], ife-2[ok306] ; Ex [p ife-2 GFP, pRF4]) Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center) Cycloheximide (Sigma-Aldrich, catalog number: C7698) Note: Cycloheximide has significant, toxic side effects. .. Potassium phosphate monobasic (KH 2 PO 4 ) (Sigma-Aldrich, catalog number: 7778-77-0) Potassium phosphate dibasic (K 2 HPO 4 ) (Sigma-Aldrich, catalog number: 7758-11-4) Sodium chloride (NaCl) (EMD Millipore, catalog number: 106404) Peptone (BD, Bacto, catalog number: 211677) Streptomycin (Sigma-Aldrich, catalog number: S6501) Agar (Sigma-Aldrich, catalog number: 05040) Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01) Calcium chloride dehydrate (CaCl 2 ⋅2H 2 O) (Sigma-Aldrich, catalog number: C5080) Magnesium sulfate (MgSO 4 ) (Sigma-Aldrich, catalog number: M7506) Nystatin stock solution (Sigma-Aldrich, catalog number: N3503) Sodium phosphate dibasic (Na 2 HPO 4 ) (Sigma-Aldrich, catalog number: 7558-79-4) Phosphate buffer (1 M; sterile, see Recipes ) Nematode growth medium (NGM) agar plates (see Recipes ) M9 buffer (see Recipes )
Transgenic Assay:Article Title: In vivo Mitophagy Monitoring in Caenorhabditis elegans to Determine Mitochondrial Homeostasis
Article Snippet: .. Greiner Petri dishes (60 x 15 mm) (Greiner Bio One International, catalog number: 628161) Microscope slides 75 x 25 x 1 mm (Marienfeld-Superior, catalog number: 10 006 12) Microscope cover glass 18 x 18 mm (Marienfeld-Superior, catalog number: 01 010 30) Use the following transgenic nematodes to monitor mitophagy: IR1631: N2; Ex003 [p myo-3 TOMM-20::Rosella; pRF4], R1284: N2; Is [p myo-3 mtGFP]; Ex011 [p lgg-1 DsRed::LGG-1; p myo-2 GFP]) Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center) 70% of EtOH Potassium dihydrogen phosphate (KH 2 PO 4 ) (EMD Millipore, catalog number: 104873) di-Potassium hydrogen phosphate (K 2 HPO 4 ) (EMD Millipore, catalog number: 137010) Sodium chloride (NaCl) (EMD Millipore, catalog number: 106404) Peptone (BD, BactoTM, catalog number: 211677) Streptomycin (Sigma-Aldrich, catalog number: S6501) Agar (Sigma-Aldrich, catalog number: 05040) Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01) Calcium chloride dehydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: C5080) Magnesium sulfate (MgSO 4 ) (Sigma-Aldrich, catalog number: M7506) Nystatin stock solution (Sigma-Aldrich, catalog number: N3503) di-Sodium hydrogen phosphate (Na 2 HPO 4 ) (EMD Millipore, catalog number: 106586) Levamisole (Sigma-Aldrich, catalog number: L9756) Paraquat (Sigma-Aldrich, catalog number: 856177) Carbonyl cyanide m-chlorophenylhydrazone (CCCP) (Sigma-Aldrich, catalog number: C2759) Dimethyl sulfoxide cell culture grade BC (DMSO) (AppliChem, catalog number: A3672,0250) Phosphate buffer (1 M; sterile, see Recipes ) Nematode growth medium (NGM) agar plates (see Recipes ) M9 buffer (see Recipes ) Levamisole (0.5 M, see Recipes ) M9-levamisole solution (20 mM solution, see Recipes ) Paraquat (0.5 M, see Recipes ) Carbonyl cyanide m-chlorophenylhydrazone (49 mM; CCCP, see Recipes ) .. UV crosslinker (Vilber Lourmat, model: BIO-LINK – BLX-E365) Zeiss AxioImager Z2 epifluorescence microscope (Zeiss, model: Zeiss AxioImager Z2) Olympus DP71 CCD camera (Olympus, model: Olympus DP71) Zeiss AxioObserver Z1 confocal microscope (Zeiss, model: Zeiss AxioObserver Z1) Dissecting stereomicroscope (Olympus, model: SMZ645) Incubators for stable temperature (AQUA ® LYTIC incubator 20 °C)
Cell Culture:Article Title: In vivo Mitophagy Monitoring in Caenorhabditis elegans to Determine Mitochondrial Homeostasis
Article Snippet: .. Greiner Petri dishes (60 x 15 mm) (Greiner Bio One International, catalog number: 628161) Microscope slides 75 x 25 x 1 mm (Marienfeld-Superior, catalog number: 10 006 12) Microscope cover glass 18 x 18 mm (Marienfeld-Superior, catalog number: 01 010 30) Use the following transgenic nematodes to monitor mitophagy: IR1631: N2; Ex003 [p myo-3 TOMM-20::Rosella; pRF4], R1284: N2; Is [p myo-3 mtGFP]; Ex011 [p lgg-1 DsRed::LGG-1; p myo-2 GFP]) Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center) 70% of EtOH Potassium dihydrogen phosphate (KH 2 PO 4 ) (EMD Millipore, catalog number: 104873) di-Potassium hydrogen phosphate (K 2 HPO 4 ) (EMD Millipore, catalog number: 137010) Sodium chloride (NaCl) (EMD Millipore, catalog number: 106404) Peptone (BD, BactoTM, catalog number: 211677) Streptomycin (Sigma-Aldrich, catalog number: S6501) Agar (Sigma-Aldrich, catalog number: 05040) Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01) Calcium chloride dehydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: C5080) Magnesium sulfate (MgSO 4 ) (Sigma-Aldrich, catalog number: M7506) Nystatin stock solution (Sigma-Aldrich, catalog number: N3503) di-Sodium hydrogen phosphate (Na 2 HPO 4 ) (EMD Millipore, catalog number: 106586) Levamisole (Sigma-Aldrich, catalog number: L9756) Paraquat (Sigma-Aldrich, catalog number: 856177) Carbonyl cyanide m-chlorophenylhydrazone (CCCP) (Sigma-Aldrich, catalog number: C2759) Dimethyl sulfoxide cell culture grade BC (DMSO) (AppliChem, catalog number: A3672,0250) Phosphate buffer (1 M; sterile, see Recipes ) Nematode growth medium (NGM) agar plates (see Recipes ) M9 buffer (see Recipes ) Levamisole (0.5 M, see Recipes ) M9-levamisole solution (20 mM solution, see Recipes ) Paraquat (0.5 M, see Recipes ) Carbonyl cyanide m-chlorophenylhydrazone (49 mM; CCCP, see Recipes ) .. UV crosslinker (Vilber Lourmat, model: BIO-LINK – BLX-E365) Zeiss AxioImager Z2 epifluorescence microscope (Zeiss, model: Zeiss AxioImager Z2) Olympus DP71 CCD camera (Olympus, model: Olympus DP71) Zeiss AxioObserver Z1 confocal microscope (Zeiss, model: Zeiss AxioObserver Z1) Dissecting stereomicroscope (Olympus, model: SMZ645) Incubators for stable temperature (AQUA ® LYTIC incubator 20 °C)
Sterility:Article Title: In vivo Mitophagy Monitoring in Caenorhabditis elegans to Determine Mitochondrial Homeostasis
Article Snippet: .. Greiner Petri dishes (60 x 15 mm) (Greiner Bio One International, catalog number: 628161) Microscope slides 75 x 25 x 1 mm (Marienfeld-Superior, catalog number: 10 006 12) Microscope cover glass 18 x 18 mm (Marienfeld-Superior, catalog number: 01 010 30) Use the following transgenic nematodes to monitor mitophagy: IR1631: N2; Ex003 [p myo-3 TOMM-20::Rosella; pRF4], R1284: N2; Is [p myo-3 mtGFP]; Ex011 [p lgg-1 DsRed::LGG-1; p myo-2 GFP]) Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center) 70% of EtOH Potassium dihydrogen phosphate (KH 2 PO 4 ) (EMD Millipore, catalog number: 104873) di-Potassium hydrogen phosphate (K 2 HPO 4 ) (EMD Millipore, catalog number: 137010) Sodium chloride (NaCl) (EMD Millipore, catalog number: 106404) Peptone (BD, BactoTM, catalog number: 211677) Streptomycin (Sigma-Aldrich, catalog number: S6501) Agar (Sigma-Aldrich, catalog number: 05040) Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01) Calcium chloride dehydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: C5080) Magnesium sulfate (MgSO 4 ) (Sigma-Aldrich, catalog number: M7506) Nystatin stock solution (Sigma-Aldrich, catalog number: N3503) di-Sodium hydrogen phosphate (Na 2 HPO 4 ) (EMD Millipore, catalog number: 106586) Levamisole (Sigma-Aldrich, catalog number: L9756) Paraquat (Sigma-Aldrich, catalog number: 856177) Carbonyl cyanide m-chlorophenylhydrazone (CCCP) (Sigma-Aldrich, catalog number: C2759) Dimethyl sulfoxide cell culture grade BC (DMSO) (AppliChem, catalog number: A3672,0250) Phosphate buffer (1 M; sterile, see Recipes ) Nematode growth medium (NGM) agar plates (see Recipes ) M9 buffer (see Recipes ) Levamisole (0.5 M, see Recipes ) M9-levamisole solution (20 mM solution, see Recipes ) Paraquat (0.5 M, see Recipes ) Carbonyl cyanide m-chlorophenylhydrazone (49 mM; CCCP, see Recipes ) .. UV crosslinker (Vilber Lourmat, model: BIO-LINK – BLX-E365) Zeiss AxioImager Z2 epifluorescence microscope (Zeiss, model: Zeiss AxioImager Z2) Olympus DP71 CCD camera (Olympus, model: Olympus DP71) Zeiss AxioObserver Z1 confocal microscope (Zeiss, model: Zeiss AxioObserver Z1) Dissecting stereomicroscope (Olympus, model: SMZ645) Incubators for stable temperature (AQUA ® LYTIC incubator 20 °C)
Article Title: Measuring Oxygen Consumption Rate in Caenorhabditis elegans
Article Snippet: In this protocol, we describe a method for the determination of oxygen consumption rates in the nematode C. elegans by using a Clark-type polarographic oxygen sensor electrode (Hansatech, King’s Lynn, England). .. Materials and Reagents list-behavior=enumerated prefix-word= mark-type=upper-alpha max-label-size=0 Consumables list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Commercial cigarette paper Polytetrafluorethylene (PTFE) membrane (provided by Hansatech, King’s Lynn, England) 15 ml tube (STARSTEDT, catalog number: 62.554.016) 1.5 ml tube (Sigma-Aldrich, catalog number: Z606340) Paper towel Greiner Petri dishes (60 x 15 mm) (Greiner Bio One, catalog number: 628161) Maintenance kit (Hansatech, King’s Lynn, England) Biological reagents list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 C. elegans strains (wild type [N2] and dct-1[tm376] ) Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center) Chemical reagents list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Distilled water Nitrogen gas provided in a tank PierceTM BCA Protein Assay Kit (Thermo Fisher Scientific, Thermo ScientificTM, catalog number: 23225) Potassium dihydrogen phosphate (KH 2 PO 4 ) (EMD Millipore, catalog number: 1048731000) K 2 HPO 4 Sodium chloride (NaCl) (EMD Millipore, catalog number: 1064041000) BactoTM peptone (BD, BactoTM, catalog number: 211677) Streptomycin (Sigma-Aldrich, catalog number: S-6501) Agar (Sigma-Aldrich, catalog number: 05040) Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01) Calcium chloride (CaCl 2 ) (Sigma-Aldrich, catalog number: C-5080) Magnesium sulfate (MgSO 4 ) (Sigma-Aldrich, catalog number: M-7506) Nystatin stock solution (Sigma-Aldrich, catalog number: N-3503) Na 2 HPO 4 (EMD Millipore, catalog number: 1065860500) Potassium chloride (KCl) (Sigma-Aldrich, catalog number: P-5405) Potassium chloride (KCl) buffer (see Recipes ) Nematode growth medium (NGM) agar plates (see Recipes ) M9 buffer (sterile; see Recipes ) Phosphate buffer (sterile; see Recipes ) Equipment list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Dissecting stereomicroscope (Olympus, model: SMZ645) Incubators for stable temperature (AQUA ® LYTIC incubator 20 °C) DW1/AD clark-type polarographic oxygen sensor (Hansatech Instruments, model: Oxygraph Plus System) Water bath at 20 °C Tabletop centrifuge (Eppendorf, model: 5424) Sonicator (Sonics & Material, model: VC 130PB) Software list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Oxygraph Plus software (Hansatech, King’s Lynn, England) Microsoft Office 2011 Excel Procedure list-behavior=enumerated prefix-word= mark-type=upper-alpha max-label-size=0 Growth and synchronization of nematode population list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Use a dissecting stereomicroscope to select L4 larvae of each strain. ..
Subculturing Assay:Article Title: Compositions suitable for treating cutaneous signs of aging
Article Snippet: B 2001, 61:106-113): Fibroblasts from human skin were cultured at 37° C. under 5% carbon dioxide in Dulbecco's Modified Eagle's Medium supplemented with 2.5% fetal calf serum (Hyclone, Logan, Utah) and 100 IU/l penicillin and 100 μg/I streptomycin. .. For subculturing the cells were detached with 0.25% trypsin/EDTA solution (Gibco) and split 1:5 and maintained in control medium in 90 mm Greiner Petri dishes. ..
Control:Article Title: Compositions suitable for treating cutaneous signs of aging
Article Snippet: B 2001, 61:106-113): Fibroblasts from human skin were cultured at 37° C. under 5% carbon dioxide in Dulbecco's Modified Eagle's Medium supplemented with 2.5% fetal calf serum (Hyclone, Logan, Utah) and 100 IU/l penicillin and 100 μg/I streptomycin. .. For subculturing the cells were detached with 0.25% trypsin/EDTA solution (Gibco) and split 1:5 and maintained in control medium in 90 mm Greiner Petri dishes. ..
Membrane:Article Title: Measuring Oxygen Consumption Rate in Caenorhabditis elegans
Article Snippet: In this protocol, we describe a method for the determination of oxygen consumption rates in the nematode C. elegans by using a Clark-type polarographic oxygen sensor electrode (Hansatech, King’s Lynn, England). .. Materials and Reagents list-behavior=enumerated prefix-word= mark-type=upper-alpha max-label-size=0 Consumables list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Commercial cigarette paper Polytetrafluorethylene (PTFE) membrane (provided by Hansatech, King’s Lynn, England) 15 ml tube (STARSTEDT, catalog number: 62.554.016) 1.5 ml tube (Sigma-Aldrich, catalog number: Z606340) Paper towel Greiner Petri dishes (60 x 15 mm) (Greiner Bio One, catalog number: 628161) Maintenance kit (Hansatech, King’s Lynn, England) Biological reagents list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 C. elegans strains (wild type [N2] and dct-1[tm376] ) Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center) Chemical reagents list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Distilled water Nitrogen gas provided in a tank PierceTM BCA Protein Assay Kit (Thermo Fisher Scientific, Thermo ScientificTM, catalog number: 23225) Potassium dihydrogen phosphate (KH 2 PO 4 ) (EMD Millipore, catalog number: 1048731000) K 2 HPO 4 Sodium chloride (NaCl) (EMD Millipore, catalog number: 1064041000) BactoTM peptone (BD, BactoTM, catalog number: 211677) Streptomycin (Sigma-Aldrich, catalog number: S-6501) Agar (Sigma-Aldrich, catalog number: 05040) Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01) Calcium chloride (CaCl 2 ) (Sigma-Aldrich, catalog number: C-5080) Magnesium sulfate (MgSO 4 ) (Sigma-Aldrich, catalog number: M-7506) Nystatin stock solution (Sigma-Aldrich, catalog number: N-3503) Na 2 HPO 4 (EMD Millipore, catalog number: 1065860500) Potassium chloride (KCl) (Sigma-Aldrich, catalog number: P-5405) Potassium chloride (KCl) buffer (see Recipes ) Nematode growth medium (NGM) agar plates (see Recipes ) M9 buffer (sterile; see Recipes ) Phosphate buffer (sterile; see Recipes ) Equipment list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Dissecting stereomicroscope (Olympus, model: SMZ645) Incubators for stable temperature (AQUA ® LYTIC incubator 20 °C) DW1/AD clark-type polarographic oxygen sensor (Hansatech Instruments, model: Oxygraph Plus System) Water bath at 20 °C Tabletop centrifuge (Eppendorf, model: 5424) Sonicator (Sonics & Material, model: VC 130PB) Software list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Oxygraph Plus software (Hansatech, King’s Lynn, England) Microsoft Office 2011 Excel Procedure list-behavior=enumerated prefix-word= mark-type=upper-alpha max-label-size=0 Growth and synchronization of nematode population list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Use a dissecting stereomicroscope to select L4 larvae of each strain. ..
Bicinchoninic Acid Protein Assay:Article Title: Measuring Oxygen Consumption Rate in Caenorhabditis elegans
Article Snippet: In this protocol, we describe a method for the determination of oxygen consumption rates in the nematode C. elegans by using a Clark-type polarographic oxygen sensor electrode (Hansatech, King’s Lynn, England). .. Materials and Reagents list-behavior=enumerated prefix-word= mark-type=upper-alpha max-label-size=0 Consumables list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Commercial cigarette paper Polytetrafluorethylene (PTFE) membrane (provided by Hansatech, King’s Lynn, England) 15 ml tube (STARSTEDT, catalog number: 62.554.016) 1.5 ml tube (Sigma-Aldrich, catalog number: Z606340) Paper towel Greiner Petri dishes (60 x 15 mm) (Greiner Bio One, catalog number: 628161) Maintenance kit (Hansatech, King’s Lynn, England) Biological reagents list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 C. elegans strains (wild type [N2] and dct-1[tm376] ) Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center) Chemical reagents list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Distilled water Nitrogen gas provided in a tank PierceTM BCA Protein Assay Kit (Thermo Fisher Scientific, Thermo ScientificTM, catalog number: 23225) Potassium dihydrogen phosphate (KH 2 PO 4 ) (EMD Millipore, catalog number: 1048731000) K 2 HPO 4 Sodium chloride (NaCl) (EMD Millipore, catalog number: 1064041000) BactoTM peptone (BD, BactoTM, catalog number: 211677) Streptomycin (Sigma-Aldrich, catalog number: S-6501) Agar (Sigma-Aldrich, catalog number: 05040) Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01) Calcium chloride (CaCl 2 ) (Sigma-Aldrich, catalog number: C-5080) Magnesium sulfate (MgSO 4 ) (Sigma-Aldrich, catalog number: M-7506) Nystatin stock solution (Sigma-Aldrich, catalog number: N-3503) Na 2 HPO 4 (EMD Millipore, catalog number: 1065860500) Potassium chloride (KCl) (Sigma-Aldrich, catalog number: P-5405) Potassium chloride (KCl) buffer (see Recipes ) Nematode growth medium (NGM) agar plates (see Recipes ) M9 buffer (sterile; see Recipes ) Phosphate buffer (sterile; see Recipes ) Equipment list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Dissecting stereomicroscope (Olympus, model: SMZ645) Incubators for stable temperature (AQUA ® LYTIC incubator 20 °C) DW1/AD clark-type polarographic oxygen sensor (Hansatech Instruments, model: Oxygraph Plus System) Water bath at 20 °C Tabletop centrifuge (Eppendorf, model: 5424) Sonicator (Sonics & Material, model: VC 130PB) Software list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Oxygraph Plus software (Hansatech, King’s Lynn, England) Microsoft Office 2011 Excel Procedure list-behavior=enumerated prefix-word= mark-type=upper-alpha max-label-size=0 Growth and synchronization of nematode population list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Use a dissecting stereomicroscope to select L4 larvae of each strain. ..
Software:Article Title: Measuring Oxygen Consumption Rate in Caenorhabditis elegans
Article Snippet: In this protocol, we describe a method for the determination of oxygen consumption rates in the nematode C. elegans by using a Clark-type polarographic oxygen sensor electrode (Hansatech, King’s Lynn, England). .. Materials and Reagents list-behavior=enumerated prefix-word= mark-type=upper-alpha max-label-size=0 Consumables list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Commercial cigarette paper Polytetrafluorethylene (PTFE) membrane (provided by Hansatech, King’s Lynn, England) 15 ml tube (STARSTEDT, catalog number: 62.554.016) 1.5 ml tube (Sigma-Aldrich, catalog number: Z606340) Paper towel Greiner Petri dishes (60 x 15 mm) (Greiner Bio One, catalog number: 628161) Maintenance kit (Hansatech, King’s Lynn, England) Biological reagents list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 C. elegans strains (wild type [N2] and dct-1[tm376] ) Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center) Chemical reagents list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Distilled water Nitrogen gas provided in a tank PierceTM BCA Protein Assay Kit (Thermo Fisher Scientific, Thermo ScientificTM, catalog number: 23225) Potassium dihydrogen phosphate (KH 2 PO 4 ) (EMD Millipore, catalog number: 1048731000) K 2 HPO 4 Sodium chloride (NaCl) (EMD Millipore, catalog number: 1064041000) BactoTM peptone (BD, BactoTM, catalog number: 211677) Streptomycin (Sigma-Aldrich, catalog number: S-6501) Agar (Sigma-Aldrich, catalog number: 05040) Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01) Calcium chloride (CaCl 2 ) (Sigma-Aldrich, catalog number: C-5080) Magnesium sulfate (MgSO 4 ) (Sigma-Aldrich, catalog number: M-7506) Nystatin stock solution (Sigma-Aldrich, catalog number: N-3503) Na 2 HPO 4 (EMD Millipore, catalog number: 1065860500) Potassium chloride (KCl) (Sigma-Aldrich, catalog number: P-5405) Potassium chloride (KCl) buffer (see Recipes ) Nematode growth medium (NGM) agar plates (see Recipes ) M9 buffer (sterile; see Recipes ) Phosphate buffer (sterile; see Recipes ) Equipment list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Dissecting stereomicroscope (Olympus, model: SMZ645) Incubators for stable temperature (AQUA ® LYTIC incubator 20 °C) DW1/AD clark-type polarographic oxygen sensor (Hansatech Instruments, model: Oxygraph Plus System) Water bath at 20 °C Tabletop centrifuge (Eppendorf, model: 5424) Sonicator (Sonics & Material, model: VC 130PB) Software list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Oxygraph Plus software (Hansatech, King’s Lynn, England) Microsoft Office 2011 Excel Procedure list-behavior=enumerated prefix-word= mark-type=upper-alpha max-label-size=0 Growth and synchronization of nematode population list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Use a dissecting stereomicroscope to select L4 larvae of each strain. ..
other:Article Title: Measuring Oxygen Consumption Rate in Caenorhabditis elegans
Article Snippet: Dissecting stereomicroscope (Olympus, model: SMZ645) Incubators for stable temperature (AQUA ® LYTIC incubator 20 °C) DW1/AD clark-type polarographic oxygen sensor (Hansatech Instruments, model: Oxygraph Plus System) Water bath at 20 °C Tabletop centrifuge (Eppendorf, model: 5424) Sonicator (Sonics & Material, model: VC 130PB)
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