Concentration Assay:Article Title: Modulation of human dorsal root ganglion neuron excitability by Nav1.7 inhibition
Article Snippet: .. The concentration-dependence for inhibition of cloned human Nav1.7 channels by AM-2099 was determined using a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line previously generated in the laboratory ( ) with human Nav1.7 channels (GenBank Accession Number X82835.1 ) expressed in a HEK293 cell line purchased from American Type Culture Collection (CAT# ATCC CRL-1573, ATCC, Manassas, VA). ..
Inhibition:Article Title: Modulation of human dorsal root ganglion neuron excitability by Nav1.7 inhibition
Article Snippet: .. The concentration-dependence for inhibition of cloned human Nav1.7 channels by AM-2099 was determined using a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line previously generated in the laboratory ( ) with human Nav1.7 channels (GenBank Accession Number X82835.1 ) expressed in a HEK293 cell line purchased from American Type Culture Collection (CAT# ATCC CRL-1573, ATCC, Manassas, VA). ..
Clone Assay:Article Title: Modulation of human dorsal root ganglion neuron excitability by Nav1.7 inhibition
Article Snippet: .. The concentration-dependence for inhibition of cloned human Nav1.7 channels by AM-2099 was determined using a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line previously generated in the laboratory ( ) with human Nav1.7 channels (GenBank Accession Number X82835.1 ) expressed in a HEK293 cell line purchased from American Type Culture Collection (CAT# ATCC CRL-1573, ATCC, Manassas, VA). ..
Automated:Article Title: Modulation of human dorsal root ganglion neuron excitability by Nav1.7 inhibition
Article Snippet: .. The concentration-dependence for inhibition of cloned human Nav1.7 channels by AM-2099 was determined using a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line previously generated in the laboratory ( ) with human Nav1.7 channels (GenBank Accession Number X82835.1 ) expressed in a HEK293 cell line purchased from American Type Culture Collection (CAT# ATCC CRL-1573, ATCC, Manassas, VA). ..
Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Patch Clamp:Article Title: Modulation of human dorsal root ganglion neuron excitability by Nav1.7 inhibition
Article Snippet: .. The concentration-dependence for inhibition of cloned human Nav1.7 channels by AM-2099 was determined using a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line previously generated in the laboratory ( ) with human Nav1.7 channels (GenBank Accession Number X82835.1 ) expressed in a HEK293 cell line purchased from American Type Culture Collection (CAT# ATCC CRL-1573, ATCC, Manassas, VA). ..
Stable Transfection:Article Title: Modulation of human dorsal root ganglion neuron excitability by Nav1.7 inhibition
Article Snippet: .. The concentration-dependence for inhibition of cloned human Nav1.7 channels by AM-2099 was determined using a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line previously generated in the laboratory ( ) with human Nav1.7 channels (GenBank Accession Number X82835.1 ) expressed in a HEK293 cell line purchased from American Type Culture Collection (CAT# ATCC CRL-1573, ATCC, Manassas, VA). ..
Generated:Article Title: Modulation of human dorsal root ganglion neuron excitability by Nav1.7 inhibition
Article Snippet: .. The concentration-dependence for inhibition of cloned human Nav1.7 channels by AM-2099 was determined using a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line previously generated in the laboratory ( ) with human Nav1.7 channels (GenBank Accession Number X82835.1 ) expressed in a HEK293 cell line purchased from American Type Culture Collection (CAT# ATCC CRL-1573, ATCC, Manassas, VA). ..
Chloramphenicol Acetyltransferase Assay:Article Title: Modulation of human dorsal root ganglion neuron excitability by Nav1.7 inhibition
Article Snippet: .. The concentration-dependence for inhibition of cloned human Nav1.7 channels by AM-2099 was determined using a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line previously generated in the laboratory ( ) with human Nav1.7 channels (GenBank Accession Number X82835.1 ) expressed in a HEK293 cell line purchased from American Type Culture Collection (CAT# ATCC CRL-1573, ATCC, Manassas, VA). ..
Virus:Article Title: Epigallocatechin gallate inhibits rabies virus glycoprotein‑mediated infection in vitro.
Article Snippet: 1 Department of Molecular Medicine, University of Padua, Padua, Italy 2 National and FAO Reference Center for Rabies, Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro, Padua, Italy 3 Department of Biology, University of Padua, Padua, Italy Abstract Rabies is one of the most lethal zoonotic disease and causes approximately 60,000 human deaths annually.. The high cost of immunoglobulin prophylaxis and the need for cold chain maintenance highlight the need for accessible antiviral strategies, particularly in resource-limited settings.. Epigallocatechin gallate (EGCG), the major polyphenolic compound of Camellia sinensis, has broad-spectrum antiviral activity.
Modification:Article Title: Epigallocatechin gallate inhibits rabies virus glycoprotein‑mediated infection in vitro.
Article Snippet: 1 Department of Molecular Medicine, University of Padua, Padua, Italy 2 National and FAO Reference Center for Rabies, Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro, Padua, Italy 3 Department of Biology, University of Padua, Padua, Italy Abstract Rabies is one of the most lethal zoonotic disease and causes approximately 60,000 human deaths annually.. The high cost of immunoglobulin prophylaxis and the need for cold chain maintenance highlight the need for accessible antiviral strategies, particularly in resource-limited settings.. Epigallocatechin gallate (EGCG), the major polyphenolic compound of Camellia sinensis, has broad-spectrum antiviral activity.
Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Incubator:
Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Control:Article Title: Isolation, Taxonomic Assignment, and Multi‐Assay Bioactivity Profiling of Sodaleptolyngbya sp. HX‐01 From Paddy Soil in Hoa Xuan
Article Snippet: .. Four human cancer cell lines, namely—KB (oral carcinoma, ATCC CCL‐17; RRID:CVCL_0372), HepG2 (hepatoblastoma; ATCC HB‐8065; RRID:CVCL_0027), A549 (lung adenocarcinoma; ATCC CCL‐185; RRID:CVCL_0023), MCF‐7 (breast adenocarcinoma; ATCC HTB‐22; RRID:CVCL_0031), and with HEK‐293 (adenovirus‐transformed, human embryonic kidney‐derived; ATCC CRL‐1573; RRID:CVCL_0045) as the nontumor control were procured from ATCC (American Type Culture Collection) were tested against extracted bioactive metabolites from isolated cyanobacterium. ..
Isolation:Article Title: Isolation, Taxonomic Assignment, and Multi‐Assay Bioactivity Profiling of Sodaleptolyngbya sp. HX‐01 From Paddy Soil in Hoa Xuan
Article Snippet: .. Four human cancer cell lines, namely—KB (oral carcinoma, ATCC CCL‐17; RRID:CVCL_0372), HepG2 (hepatoblastoma; ATCC HB‐8065; RRID:CVCL_0027), A549 (lung adenocarcinoma; ATCC CCL‐185; RRID:CVCL_0023), MCF‐7 (breast adenocarcinoma; ATCC HTB‐22; RRID:CVCL_0031), and with HEK‐293 (adenovirus‐transformed, human embryonic kidney‐derived; ATCC CRL‐1573; RRID:CVCL_0045) as the nontumor control were procured from ATCC (American Type Culture Collection) were tested against extracted bioactive metabolites from isolated cyanobacterium. ..
Saline:Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Staining:Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Transfection:Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Cell Culture:Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Cell Counting:Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Polymerase Chain Reaction:Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Sterility:Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Qubit:Article Title: Protocol for Designing and Interpreting Minigene Assays to Validate Candidate Splice‐Altering Variants Using Long‐read Sequencing
Article Snippet: .. Complete growth medium (see recipe) Mammalian cell line to transfect (e.g., HEK‐293, ATCC #CRL‐1573) Phosphate‐buffered saline (PBS), pH 7.4 (Gibco, cat. no. 10010023) Trypsin‐EDTA (0.25%), phenol red (Thermo Fisher, cat. no. 25200072) Trypan Blue Stain (Invitrogen, cat no. T10282 ) Lipofectamine 3000 transfection reagent (Invitrogen, cat. no. L3000008) Opti‐MEM reduced serum medium (Gibco, cat. no. 31985062) Minigene plasmids as designed in Basic Protocol Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) Cycloheximide (Sigma‐Aldrich, cat. no. C4859) RNeasy Mini Kit (Qiagen, cat. no. 74106) SuperScript III First‐Strand Synthesis SuperMix (Invitrogen, cat. no. 18080400) Labculture Class II Type A2 Biological Safety Cabinet (Esco Lifesciences) Julabo Eco Temp TW12 water bath (Sigma‐Aldrich) Cell Culture Flask, 250 mL, 75 cm2, Ps, Red Filter Screw Cap, Clear (Greiner Bio‐One, cat. no. 658175) Tube, 15 mL, Pp, 17/120 Mm, Conical Bottom (Greiner Bio‐One, cat. no. 188271) Heracel VIOS 160i CO 2 Incubator (Thermo Fisher Scientific) Cell Culture Multiwell Plate, 6 Well, Ps, Clear (Greiner Bio‐One, cat. no. 657160) Countess Cell Counting Chamber Slides (Invitrogen, cat. no. C10228 ) Automated Cell Counter ‐ Countess II FL (Life Technologies) Microplate, 96 Well, Ps, F‐Bottom, Clear (Greiner Bio‐One, cat. no. 655101) Mastercycler Nexus Gradient PCR thermocycler (Eppendorf) Centrifuge 5810 R (Eppendorf) Tube 1.5 mL Sterile (SSI Bio, cat. no. 1210‐S0) Reax top test tube shaker (vortex) (Heidolph) MIKRO 220 Microcentrifuge (Hettich) Qubit 4 Fluorometer (Invitrogen) ..
Binding Assay:
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