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Millipore exonuclease digestion
Exonuclease Digestion, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/exonuclease digestion/product/Millipore
Average 92 stars, based on 1 article reviews
Price from $9.99 to $1999.99
exonuclease digestion - by Bioz Stars, 2021-07
92/100 stars

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Recombinant:

Article Title: Distribution of the Galβ1-4Gal Epitope among Birds: Species-Specific Loss of the Glycan Structure in Chicken and Its Relatives
Article Snippet: .. The N -glycans were digested with various exo-glycosidases under the following conditions: 2 mU of α2,3-specific neuraminidase from Macrobdella decora (Recombinant, Calbiochem) in 20 µl of 50 mM ammonium acetate buffer (pH 6.0), at 37°C for 24 h; 10 mU of α-galactosidase from green coffee bean (Calbiochem) in 20 µl of 50 mM ammonium acetate buffer (pH 6.0), at 37°C for 48 h; and 3 mU of β1,4-galactosidase from Streptococcus pneumoniae (Recombinant, Calbiochem) in 20 µl of 50 mM ammonium acetate buffer (pH 6.0), at 37°C for 24 h. Permethylation of the released N -glycans, and subsequent MALDI-MS and MS/MS analyses on a MALDI TOF/TOF (ABI 4700 Proteomic Analyzer) were performed essentially as described previously . .. Additional nano electrospray ionization (nanoESI)-MSn analyses were performed on an LTQ-Orbitrap XL hybrid FT mass spectrometer (Thermo Scientific).

Tandem Mass Spectroscopy:

Article Title: Distribution of the Galβ1-4Gal Epitope among Birds: Species-Specific Loss of the Glycan Structure in Chicken and Its Relatives
Article Snippet: .. The N -glycans were digested with various exo-glycosidases under the following conditions: 2 mU of α2,3-specific neuraminidase from Macrobdella decora (Recombinant, Calbiochem) in 20 µl of 50 mM ammonium acetate buffer (pH 6.0), at 37°C for 24 h; 10 mU of α-galactosidase from green coffee bean (Calbiochem) in 20 µl of 50 mM ammonium acetate buffer (pH 6.0), at 37°C for 48 h; and 3 mU of β1,4-galactosidase from Streptococcus pneumoniae (Recombinant, Calbiochem) in 20 µl of 50 mM ammonium acetate buffer (pH 6.0), at 37°C for 24 h. Permethylation of the released N -glycans, and subsequent MALDI-MS and MS/MS analyses on a MALDI TOF/TOF (ABI 4700 Proteomic Analyzer) were performed essentially as described previously . .. Additional nano electrospray ionization (nanoESI)-MSn analyses were performed on an LTQ-Orbitrap XL hybrid FT mass spectrometer (Thermo Scientific).

Activity Assay:

Article Title: Enzymatic Mechanism for Arabinan Degradation and Transport in the Thermophilic Bacterium Caldanaerobius polysaccharolyticus
Article Snippet: To facilitate subsequent ligation into the pET46 Ek/LIC vector, the forward primers were engineered to incorporate a 5′-GACGACGACAAG extension and the reverse primers were designed to include a 5′-GAGGAGAAGCCCGGT extension. .. The resultant amplicons of the genes were treated with the exonuclease activity of T4 DNA polymerase and annealed with the pET46 Ek/LIC vector using the Ek/LIC cloning kit (Novagen). .. The LIC mixture was introduced into E. coli JM109 competent cells by electroporation using a Gene Pulser Xcell from Bio-Rad (Hercules, CA).

Article Title: Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly
Article Snippet: .. For exonuclease activity assay and dsDNA binding assay, His6 -tagged AtDECP was cloned into a pET-15(b) plasmid (Novagen), expressed in E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies), and purified using four step purification; affinity chromatography (HiTrap chelating column, GE Healthcare), size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare), ion exchange chromatography (HiTrap Q column, GE Healthcare), and size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare). .. Crystals of AtDECP were grown at 277 K by the hanging-drop vapor diffusion method from 10 mg ml−1 protein solution in buffer (50 mM NaCl, 3 mM NaN3 , 0.3 mM TCEP, 5 mM Bis-Tris pH 6.0) mixed with an equal amount of well solution containing sodium citrate, polyethylene glycol 2000, and HEPES pH 8.0.

Article Title: Mapping Viral DNA Specificity to the Central Region of Integrase by Using Functional Human Immunodeficiency Virus Type 1/Visna Virus Chimeric Proteins
Article Snippet: All oligodeoxynucleotides used as assay substrates were gel purified following synthesis and again after being 5′-end labeled with [γ-32 P]ATP by T4 polynucleotide kinase, as described previously ( ). .. Sequence-specific markers for gel analysis were produced by the 3′-to-5′ exonuclease activity of snake venom phosphodiesterase (Sigma, St. Louis, Mo.) on 5′-radiolabeled oligonucleotides, as described previously ( ). .. Double-stranded DNA substrates were prepared by annealing the labeled strand with a fourfold excess of unlabeled complementary oligonucleotide, and the disintegration assay substrate was prepared and then purified on a native 15% polyacrylamide gel, both as described previously ( ).

Article Title: Two New Xylanases with Different Substrate Specificities from the Human Gut Bacterium Bacteroides intestinalis DSM 17393
Article Snippet: The coding sequences for these three genes were amplified by PCR using the PicoMaxx high-fidelity PCR mix from Agilent, and the resulting amplicons were purified using the Wizard DNA purification kit (Promega, Madison, WI). .. The purified amplicons were digested using the exonuclease activity of T4 DNA polymerase, annealed with a similarly digested pET-46b vector (EMD Chemicals, Darmstadt, Germany), and introduced into E. coli XL10 Gold competent cells by chemical transformation. .. Individual colonies were picked and cultured overnight in lysogeny broth (LB) supplemented with ampicillin (100 μg/ml), and plasmid DNA was purified using a Plasmid Minikit from Qiagen (Valencia, CA).

Plasmid Preparation:

Article Title: Enzymatic Mechanism for Arabinan Degradation and Transport in the Thermophilic Bacterium Caldanaerobius polysaccharolyticus
Article Snippet: To facilitate subsequent ligation into the pET46 Ek/LIC vector, the forward primers were engineered to incorporate a 5′-GACGACGACAAG extension and the reverse primers were designed to include a 5′-GAGGAGAAGCCCGGT extension. .. The resultant amplicons of the genes were treated with the exonuclease activity of T4 DNA polymerase and annealed with the pET46 Ek/LIC vector using the Ek/LIC cloning kit (Novagen). .. The LIC mixture was introduced into E. coli JM109 competent cells by electroporation using a Gene Pulser Xcell from Bio-Rad (Hercules, CA).

Article Title: Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly
Article Snippet: .. For exonuclease activity assay and dsDNA binding assay, His6 -tagged AtDECP was cloned into a pET-15(b) plasmid (Novagen), expressed in E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies), and purified using four step purification; affinity chromatography (HiTrap chelating column, GE Healthcare), size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare), ion exchange chromatography (HiTrap Q column, GE Healthcare), and size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare). .. Crystals of AtDECP were grown at 277 K by the hanging-drop vapor diffusion method from 10 mg ml−1 protein solution in buffer (50 mM NaCl, 3 mM NaN3 , 0.3 mM TCEP, 5 mM Bis-Tris pH 6.0) mixed with an equal amount of well solution containing sodium citrate, polyethylene glycol 2000, and HEPES pH 8.0.

Article Title: MutL sliding clamps coordinate exonuclease-independent Escherichia coli mismatch repair
Article Snippet: EcExoVII and EcSSB (unlabeled) were purchased from New England Biolabs or Thermo Fisher Scientific, respectively. .. The E. coli uvrD , ssb , and exoI(xonA) genes were amplified by PCR (Supplementary Table ), digested with XbaI , and XhoI (for UvrD ), NdeI and EcoRI (for SSB ) or NdeI and BamHI (for ExoI ), and inserted into pET-29a (Novagen) bacterial expression plasmid. .. Hexa-histidine (his6 ) and Formylglycine Generating Enzyme (FGE) recognition hexa-amino acid sequence (LCTPSR; ald6 ) were introduced onto the C-terminus of EcUvrD and EcExoI proteins.

Article Title: Two New Xylanases with Different Substrate Specificities from the Human Gut Bacterium Bacteroides intestinalis DSM 17393
Article Snippet: The coding sequences for these three genes were amplified by PCR using the PicoMaxx high-fidelity PCR mix from Agilent, and the resulting amplicons were purified using the Wizard DNA purification kit (Promega, Madison, WI). .. The purified amplicons were digested using the exonuclease activity of T4 DNA polymerase, annealed with a similarly digested pET-46b vector (EMD Chemicals, Darmstadt, Germany), and introduced into E. coli XL10 Gold competent cells by chemical transformation. .. Individual colonies were picked and cultured overnight in lysogeny broth (LB) supplemented with ampicillin (100 μg/ml), and plasmid DNA was purified using a Plasmid Minikit from Qiagen (Valencia, CA).

Clone Assay:

Article Title: Enzymatic Mechanism for Arabinan Degradation and Transport in the Thermophilic Bacterium Caldanaerobius polysaccharolyticus
Article Snippet: To facilitate subsequent ligation into the pET46 Ek/LIC vector, the forward primers were engineered to incorporate a 5′-GACGACGACAAG extension and the reverse primers were designed to include a 5′-GAGGAGAAGCCCGGT extension. .. The resultant amplicons of the genes were treated with the exonuclease activity of T4 DNA polymerase and annealed with the pET46 Ek/LIC vector using the Ek/LIC cloning kit (Novagen). .. The LIC mixture was introduced into E. coli JM109 competent cells by electroporation using a Gene Pulser Xcell from Bio-Rad (Hercules, CA).

Article Title: Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly
Article Snippet: .. For exonuclease activity assay and dsDNA binding assay, His6 -tagged AtDECP was cloned into a pET-15(b) plasmid (Novagen), expressed in E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies), and purified using four step purification; affinity chromatography (HiTrap chelating column, GE Healthcare), size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare), ion exchange chromatography (HiTrap Q column, GE Healthcare), and size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare). .. Crystals of AtDECP were grown at 277 K by the hanging-drop vapor diffusion method from 10 mg ml−1 protein solution in buffer (50 mM NaCl, 3 mM NaN3 , 0.3 mM TCEP, 5 mM Bis-Tris pH 6.0) mixed with an equal amount of well solution containing sodium citrate, polyethylene glycol 2000, and HEPES pH 8.0.

other:

Article Title: Induced Expression of Drug Metabolizing Enzymes by Preventive Agents: Role of the Antioxidant Response Element
Article Snippet: 5,6 BF, DAS, EXO and PB were obtained from Sigma Chemical Co. (St. Louis, MO).

Binding Assay:

Article Title: Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly
Article Snippet: .. For exonuclease activity assay and dsDNA binding assay, His6 -tagged AtDECP was cloned into a pET-15(b) plasmid (Novagen), expressed in E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies), and purified using four step purification; affinity chromatography (HiTrap chelating column, GE Healthcare), size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare), ion exchange chromatography (HiTrap Q column, GE Healthcare), and size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare). .. Crystals of AtDECP were grown at 277 K by the hanging-drop vapor diffusion method from 10 mg ml−1 protein solution in buffer (50 mM NaCl, 3 mM NaN3 , 0.3 mM TCEP, 5 mM Bis-Tris pH 6.0) mixed with an equal amount of well solution containing sodium citrate, polyethylene glycol 2000, and HEPES pH 8.0.

Positron Emission Tomography:

Article Title: Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly
Article Snippet: .. For exonuclease activity assay and dsDNA binding assay, His6 -tagged AtDECP was cloned into a pET-15(b) plasmid (Novagen), expressed in E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies), and purified using four step purification; affinity chromatography (HiTrap chelating column, GE Healthcare), size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare), ion exchange chromatography (HiTrap Q column, GE Healthcare), and size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare). .. Crystals of AtDECP were grown at 277 K by the hanging-drop vapor diffusion method from 10 mg ml−1 protein solution in buffer (50 mM NaCl, 3 mM NaN3 , 0.3 mM TCEP, 5 mM Bis-Tris pH 6.0) mixed with an equal amount of well solution containing sodium citrate, polyethylene glycol 2000, and HEPES pH 8.0.

Article Title: MutL sliding clamps coordinate exonuclease-independent Escherichia coli mismatch repair
Article Snippet: EcExoVII and EcSSB (unlabeled) were purchased from New England Biolabs or Thermo Fisher Scientific, respectively. .. The E. coli uvrD , ssb , and exoI(xonA) genes were amplified by PCR (Supplementary Table ), digested with XbaI , and XhoI (for UvrD ), NdeI and EcoRI (for SSB ) or NdeI and BamHI (for ExoI ), and inserted into pET-29a (Novagen) bacterial expression plasmid. .. Hexa-histidine (his6 ) and Formylglycine Generating Enzyme (FGE) recognition hexa-amino acid sequence (LCTPSR; ald6 ) were introduced onto the C-terminus of EcUvrD and EcExoI proteins.

Article Title: Two New Xylanases with Different Substrate Specificities from the Human Gut Bacterium Bacteroides intestinalis DSM 17393
Article Snippet: The coding sequences for these three genes were amplified by PCR using the PicoMaxx high-fidelity PCR mix from Agilent, and the resulting amplicons were purified using the Wizard DNA purification kit (Promega, Madison, WI). .. The purified amplicons were digested using the exonuclease activity of T4 DNA polymerase, annealed with a similarly digested pET-46b vector (EMD Chemicals, Darmstadt, Germany), and introduced into E. coli XL10 Gold competent cells by chemical transformation. .. Individual colonies were picked and cultured overnight in lysogeny broth (LB) supplemented with ampicillin (100 μg/ml), and plasmid DNA was purified using a Plasmid Minikit from Qiagen (Valencia, CA).

Purification:

Article Title: Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly
Article Snippet: .. For exonuclease activity assay and dsDNA binding assay, His6 -tagged AtDECP was cloned into a pET-15(b) plasmid (Novagen), expressed in E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies), and purified using four step purification; affinity chromatography (HiTrap chelating column, GE Healthcare), size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare), ion exchange chromatography (HiTrap Q column, GE Healthcare), and size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare). .. Crystals of AtDECP were grown at 277 K by the hanging-drop vapor diffusion method from 10 mg ml−1 protein solution in buffer (50 mM NaCl, 3 mM NaN3 , 0.3 mM TCEP, 5 mM Bis-Tris pH 6.0) mixed with an equal amount of well solution containing sodium citrate, polyethylene glycol 2000, and HEPES pH 8.0.

Article Title: Two New Xylanases with Different Substrate Specificities from the Human Gut Bacterium Bacteroides intestinalis DSM 17393
Article Snippet: The coding sequences for these three genes were amplified by PCR using the PicoMaxx high-fidelity PCR mix from Agilent, and the resulting amplicons were purified using the Wizard DNA purification kit (Promega, Madison, WI). .. The purified amplicons were digested using the exonuclease activity of T4 DNA polymerase, annealed with a similarly digested pET-46b vector (EMD Chemicals, Darmstadt, Germany), and introduced into E. coli XL10 Gold competent cells by chemical transformation. .. Individual colonies were picked and cultured overnight in lysogeny broth (LB) supplemented with ampicillin (100 μg/ml), and plasmid DNA was purified using a Plasmid Minikit from Qiagen (Valencia, CA).

Affinity Chromatography:

Article Title: Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly
Article Snippet: .. For exonuclease activity assay and dsDNA binding assay, His6 -tagged AtDECP was cloned into a pET-15(b) plasmid (Novagen), expressed in E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies), and purified using four step purification; affinity chromatography (HiTrap chelating column, GE Healthcare), size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare), ion exchange chromatography (HiTrap Q column, GE Healthcare), and size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare). .. Crystals of AtDECP were grown at 277 K by the hanging-drop vapor diffusion method from 10 mg ml−1 protein solution in buffer (50 mM NaCl, 3 mM NaN3 , 0.3 mM TCEP, 5 mM Bis-Tris pH 6.0) mixed with an equal amount of well solution containing sodium citrate, polyethylene glycol 2000, and HEPES pH 8.0.

Size-exclusion Chromatography:

Article Title: Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly
Article Snippet: .. For exonuclease activity assay and dsDNA binding assay, His6 -tagged AtDECP was cloned into a pET-15(b) plasmid (Novagen), expressed in E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies), and purified using four step purification; affinity chromatography (HiTrap chelating column, GE Healthcare), size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare), ion exchange chromatography (HiTrap Q column, GE Healthcare), and size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare). .. Crystals of AtDECP were grown at 277 K by the hanging-drop vapor diffusion method from 10 mg ml−1 protein solution in buffer (50 mM NaCl, 3 mM NaN3 , 0.3 mM TCEP, 5 mM Bis-Tris pH 6.0) mixed with an equal amount of well solution containing sodium citrate, polyethylene glycol 2000, and HEPES pH 8.0.

Ion Exchange Chromatography:

Article Title: Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly
Article Snippet: .. For exonuclease activity assay and dsDNA binding assay, His6 -tagged AtDECP was cloned into a pET-15(b) plasmid (Novagen), expressed in E. coli BL21-CodonPlus (DE3)-RIPL cells (Agilent Technologies), and purified using four step purification; affinity chromatography (HiTrap chelating column, GE Healthcare), size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare), ion exchange chromatography (HiTrap Q column, GE Healthcare), and size-exclusion chromatography (Superdex200 16/600 prep grade, GE Healthcare). .. Crystals of AtDECP were grown at 277 K by the hanging-drop vapor diffusion method from 10 mg ml−1 protein solution in buffer (50 mM NaCl, 3 mM NaN3 , 0.3 mM TCEP, 5 mM Bis-Tris pH 6.0) mixed with an equal amount of well solution containing sodium citrate, polyethylene glycol 2000, and HEPES pH 8.0.

Amplification:

Article Title: MutL sliding clamps coordinate exonuclease-independent Escherichia coli mismatch repair
Article Snippet: EcExoVII and EcSSB (unlabeled) were purchased from New England Biolabs or Thermo Fisher Scientific, respectively. .. The E. coli uvrD , ssb , and exoI(xonA) genes were amplified by PCR (Supplementary Table ), digested with XbaI , and XhoI (for UvrD ), NdeI and EcoRI (for SSB ) or NdeI and BamHI (for ExoI ), and inserted into pET-29a (Novagen) bacterial expression plasmid. .. Hexa-histidine (his6 ) and Formylglycine Generating Enzyme (FGE) recognition hexa-amino acid sequence (LCTPSR; ald6 ) were introduced onto the C-terminus of EcUvrD and EcExoI proteins.

Polymerase Chain Reaction:

Article Title: MutL sliding clamps coordinate exonuclease-independent Escherichia coli mismatch repair
Article Snippet: EcExoVII and EcSSB (unlabeled) were purchased from New England Biolabs or Thermo Fisher Scientific, respectively. .. The E. coli uvrD , ssb , and exoI(xonA) genes were amplified by PCR (Supplementary Table ), digested with XbaI , and XhoI (for UvrD ), NdeI and EcoRI (for SSB ) or NdeI and BamHI (for ExoI ), and inserted into pET-29a (Novagen) bacterial expression plasmid. .. Hexa-histidine (his6 ) and Formylglycine Generating Enzyme (FGE) recognition hexa-amino acid sequence (LCTPSR; ald6 ) were introduced onto the C-terminus of EcUvrD and EcExoI proteins.

Expressing:

Article Title: MutL sliding clamps coordinate exonuclease-independent Escherichia coli mismatch repair
Article Snippet: EcExoVII and EcSSB (unlabeled) were purchased from New England Biolabs or Thermo Fisher Scientific, respectively. .. The E. coli uvrD , ssb , and exoI(xonA) genes were amplified by PCR (Supplementary Table ), digested with XbaI , and XhoI (for UvrD ), NdeI and EcoRI (for SSB ) or NdeI and BamHI (for ExoI ), and inserted into pET-29a (Novagen) bacterial expression plasmid. .. Hexa-histidine (his6 ) and Formylglycine Generating Enzyme (FGE) recognition hexa-amino acid sequence (LCTPSR; ald6 ) were introduced onto the C-terminus of EcUvrD and EcExoI proteins.

Sequencing:

Article Title: Mapping Viral DNA Specificity to the Central Region of Integrase by Using Functional Human Immunodeficiency Virus Type 1/Visna Virus Chimeric Proteins
Article Snippet: All oligodeoxynucleotides used as assay substrates were gel purified following synthesis and again after being 5′-end labeled with [γ-32 P]ATP by T4 polynucleotide kinase, as described previously ( ). .. Sequence-specific markers for gel analysis were produced by the 3′-to-5′ exonuclease activity of snake venom phosphodiesterase (Sigma, St. Louis, Mo.) on 5′-radiolabeled oligonucleotides, as described previously ( ). .. Double-stranded DNA substrates were prepared by annealing the labeled strand with a fourfold excess of unlabeled complementary oligonucleotide, and the disintegration assay substrate was prepared and then purified on a native 15% polyacrylamide gel, both as described previously ( ).

Produced:

Article Title: Mapping Viral DNA Specificity to the Central Region of Integrase by Using Functional Human Immunodeficiency Virus Type 1/Visna Virus Chimeric Proteins
Article Snippet: All oligodeoxynucleotides used as assay substrates were gel purified following synthesis and again after being 5′-end labeled with [γ-32 P]ATP by T4 polynucleotide kinase, as described previously ( ). .. Sequence-specific markers for gel analysis were produced by the 3′-to-5′ exonuclease activity of snake venom phosphodiesterase (Sigma, St. Louis, Mo.) on 5′-radiolabeled oligonucleotides, as described previously ( ). .. Double-stranded DNA substrates were prepared by annealing the labeled strand with a fourfold excess of unlabeled complementary oligonucleotide, and the disintegration assay substrate was prepared and then purified on a native 15% polyacrylamide gel, both as described previously ( ).

Transformation Assay:

Article Title: Two New Xylanases with Different Substrate Specificities from the Human Gut Bacterium Bacteroides intestinalis DSM 17393
Article Snippet: The coding sequences for these three genes were amplified by PCR using the PicoMaxx high-fidelity PCR mix from Agilent, and the resulting amplicons were purified using the Wizard DNA purification kit (Promega, Madison, WI). .. The purified amplicons were digested using the exonuclease activity of T4 DNA polymerase, annealed with a similarly digested pET-46b vector (EMD Chemicals, Darmstadt, Germany), and introduced into E. coli XL10 Gold competent cells by chemical transformation. .. Individual colonies were picked and cultured overnight in lysogeny broth (LB) supplemented with ampicillin (100 μg/ml), and plasmid DNA was purified using a Plasmid Minikit from Qiagen (Valencia, CA).

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