Irradiation:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Amplification:Article Title: Puerarin alleviates burn-related procedural pain mediated by P2X3 receptors
Article Snippet: .. It was reverse-transcribed into complementary DNA (cDNA) with Revert Aid First Strand cDNA Synthesis Kit (Fermentas Co.). cDNA was amplified in 50 μl of polymerase chain reaction (PCR) mixture (TIANGEN Co.). ..
Fluorescence:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Labeling:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
SYBR Green Assay:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Polymerase Chain Reaction:Article Title: Climate Is Not All: Evidence From Phylogeography of Rhodiola fastigiata (Crassulaceae) and Comparison to Its Closest Relatives
Article Snippet: .. Polymerase Chain Reaction (PCR) mixture were 20 μl with 2 μl 10× buffer, 0.5 μl of each primer, 0.4 μl of dNTP mixture, 1 U of Taq polymerase (TianGen Biotech, Xi’an, China) and 1 μl template genomic DNA. .. The PCR cycling programs followed .
Article Title: Correlation of MDR1 gene polymorphisms with anesthetic effect of sevoflurane–remifentanil following pediatric tonsillectomy
Article Snippet: .. The whole reaction system was 20 μL with extracted DNA of 1 μL, forward and reverse primer (10 pmol/mL) of 1 μL for each polymerase chain reaction (PCR) mixture of 10 μL, and deionized water of 7 μL (First Strand cDNA Synthesis Kit No., KP202-03; Tiangen Biotech Co. Ltd, Beijing, China). ..
Article Title: Puerarin alleviates burn-related procedural pain mediated by P2X3 receptors
Article Snippet: .. It was reverse-transcribed into complementary DNA (cDNA) with Revert Aid First Strand cDNA Synthesis Kit (Fermentas Co.). cDNA was amplified in 50 μl of polymerase chain reaction (PCR) mixture (TIANGEN Co.). ..
Article Title: Target-site and non-target-site based resistance to the herbicide tribenuron-methyl in flixweed (Descurainia sophia L.)
Article Snippet: .. The 25 μL polymerase chain reaction (PCR) mixture consisted of 100 ng of genomic DNA, 0.5 μL (25 μM) of each primer and 12.5 μL of 2 × Taq PCR Master Mix and 10.5 μL of ddH2 O (Tiangen, Beijing, China). ..
Article Title: Molecular cloning, characterization and expression of the energy homeostasis-associated gene in piglet *
Article Snippet: .. The polymerase chain reaction (PCR) mixture (25 μl total volume) contained 12.5 μl of 2× Taq PCR master mix (Tiangen, Beijing, China), 8.5 μl of ddH2 O, 2 μl of mixed cDNA, and l μl (10 nmol/L) of each primer. .. The PCR reaction was performed using Eppendorf Mastercycler (Eppendorf, Hamburg, Germany) with the following parameters: initial denaturation at 94 °C for 5 min; 35 cycles of denaturation at 94 °C for 60 s, annealing at 55 °C for 60 s, and elongation at 72 °C for 60 s; and final elongation at 72 °C for 10 min. PCR products (4 μl) were identified by electrophoresis at 12 V/cm for 50 min on a 1% (0.01 g/ml) agarose gel.
Article Title: Significant Association Between CAV1 Variant rs3807989 on 7p31 and Atrial Fibrillation in a Chinese Han Population
Article Snippet: .. A total of 25 μL of polymerase chain reaction (PCR) mixture for genotyping contained 1.5 mmol/L of Mg2+ , 0.2 mmol/L of dNTPs, 0.5 μmol/L of each primer, 25 ng of human genomic DNA template, 5 μmol/L of SYTO 9 green fluorescent, and 0.15 U of Taq DNA polymerase (TIANGEN, Beijing, China). .. PCR was performed on an ABI 9700 System (Applied Biosystems, Foster City, CA) with a thermal profile of 95°C for 5 minutes, 40 cycles of 95°C for 10 seconds, 59.4°C or other appropriate annealing temperatures for 10 seconds and 72°C for 15 seconds, and 72°C for 10 minutes.
Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Reverse Transcription Polymerase Chain Reaction:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Real-time Polymerase Chain Reaction:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Plasmid Preparation:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
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