competent top10 e coli  (Thermo Fisher)


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    Structured Review

    Thermo Fisher competent top10 e coli
    Competent Top10 E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/competent top10 e coli/product/Thermo Fisher
    Average 99 stars, based on 25 article reviews
    Price from $9.99 to $1999.99
    competent top10 e coli - by Bioz Stars, 2020-01
    99/100 stars

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    Related Articles

    Transduction:

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA). .. Phage 80α was used to transduce the constructs from RN4220 into all other S. aureus strains used in this study.

    Clone Assay:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: Paragraph title: Cloning and sequencing of A . franciscana type 2 Hsp40 cDNA ... The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada).

    Article Title: Human Bocavirus Can Be Cultured in Differentiated Human Airway Epithelial Cells ▿
    Article Snippet: .. Purified PCR products were cloned into the pCR2.1-TOPO TA vector (Invitrogen) and transformed in chemically competent TOP10 Escherichia coli (Invitrogen), according to manufacturer protocol. .. Transformants were directly analyzed via colony PCR with T7 and M13RP primers.

    Article Title: Functional Aspects of Intrahepatic Hepatitis B Virus-specific T Cells Induced by Therapeutic DNA Vaccination
    Article Snippet: The FLuc2-gene (Promega, Madison, WI) cloned into pVAX1 has been described previously. .. Plasmids were grown in competent TOP10 Escherichia coli (Life Technologies, Carlsbad, CA) and purified using Qiagen EndoFree Plasmid purification Kit according to the manufacturer's instructions (Qiagen, Hilden, Germany).

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: Paragraph title: ccrAB cloning. ... PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA).

    Amplification:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: Primers containing the restriction enzyme sites, BamH1 and HindIII, were designed based on sequence comparisons (Integrated DNA Technologies, Coralville, IA, USA) ( ) and used for the amplification of full length A . franciscana type 2 Hsp40 cDNA from 5 h cysts. .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada).

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: Primers containing the restriction enzyme sites, BamH1 and HindIII, were designed based on sequence comparisons (Integrated DNA Technologies, Coralville, IA, USA) ( ) and used for the amplification of full length A . franciscana type 2 Hsp40 cDNA from 5 h cysts. .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada).

    Article Title: Human Bocavirus Can Be Cultured in Differentiated Human Airway Epithelial Cells ▿
    Article Snippet: RT was performed with the JZH-OligodT primer (5′-GCTATCATCACAATGGACTTTTTTTTTTTTTTTTTTV-3′), and PCR amplification was performed with the JZH-Nested adaptor primer (5′-GCTATCATCACAATGGAC-3′) and a gene-specific primer (Table ). .. Purified PCR products were cloned into the pCR2.1-TOPO TA vector (Invitrogen) and transformed in chemically competent TOP10 Escherichia coli (Invitrogen), according to manufacturer protocol.

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: ccrAB was PCR amplified from the genomic DNA of S. aureus strains N315, MW2, C98-370, C99-529, J28, J35, and J52 and Staphylococcus epidermidis strains SE5, SE7, SE50, and SE63 with primers ccuprev and ccdwnfor using the Taq PCR Master Mix kit (QIAGEN, Valencia CA.). .. PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA).

    Synthesized:

    Article Title: Efficient Site-Directed Saturation Mutagenesis Using Degenerate Oligonucleotides
    Article Snippet: Reactions were cooled on ice and digested with 5 units of Dpn I for 1 h at 37ºC to cleave methylated and hemimethylated parental DNA, but not the newly synthesized mutant DNA molecules. .. Reactions were used to transform 50 μL of chemically competent TOP10 E. coli (Invitrogen, Carlsbad, CA).

    TA Cloning:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Construct:

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA). .. A staphylococcal origin of replication was introduced by cloning plasmid pRN5543 into the unique XbaI site on PCR 2.1-TOPO, and the constructs were moved into S. aureus RN4220 by electroporation ( ).

    Electrophoresis:

    Article Title: Human Bocavirus Can Be Cultured in Differentiated Human Airway Epithelial Cells ▿
    Article Snippet: The PCR products were excised after agarose electrophoresis and purified with the NucleoSpin Extract II kit (Machery-Nagel) according to the manufacturer's protocol. .. Purified PCR products were cloned into the pCR2.1-TOPO TA vector (Invitrogen) and transformed in chemically competent TOP10 Escherichia coli (Invitrogen), according to manufacturer protocol.

    Incubation:

    Article Title: Efficient Site-Directed Saturation Mutagenesis Using Degenerate Oligonucleotides
    Article Snippet: Reactions were thermal cycled: 95ºC for 2 min, followed by 16 cycles of 95ºC for 30 sec, 55ºC for 1 min, and 68ºC for 10 min, then a final incubation of 68ºC for 10 min. .. Reactions were used to transform 50 μL of chemically competent TOP10 E. coli (Invitrogen, Carlsbad, CA).

    Transformation Assay:

    Article Title: Human Bocavirus Can Be Cultured in Differentiated Human Airway Epithelial Cells ▿
    Article Snippet: .. Purified PCR products were cloned into the pCR2.1-TOPO TA vector (Invitrogen) and transformed in chemically competent TOP10 Escherichia coli (Invitrogen), according to manufacturer protocol. .. Transformants were directly analyzed via colony PCR with T7 and M13RP primers.

    Article Title: Efficient Site-Directed Saturation Mutagenesis Using Degenerate Oligonucleotides
    Article Snippet: Reactions were used to transform 50 μL of chemically competent TOP10 E. coli (Invitrogen, Carlsbad, CA). .. Cells were heat-shocked at 42ºC for 30 sec, then placed on ice again for 1 2 min. Next, 250 μL of SOC media (Sigma Chemical, St. Louis, MO) was added, and transformed cells incubated at 37°C with shaking for 1 h. Finally, 100 150 μL was plated onto LB agar plates containing ampicillin (100μg/mL) and incubated overnight at 37ºC.

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: .. PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA). .. A staphylococcal origin of replication was introduced by cloning plasmid pRN5543 into the unique XbaI site on PCR 2.1-TOPO, and the constructs were moved into S. aureus RN4220 by electroporation ( ).

    Electroporation:

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA). .. A staphylococcal origin of replication was introduced by cloning plasmid pRN5543 into the unique XbaI site on PCR 2.1-TOPO, and the constructs were moved into S. aureus RN4220 by electroporation ( ).

    Transfection:

    Article Title: A CLAG3 Mutation in an Amphipathic Transmembrane Domain Alters Malaria Parasite Nutrient Channels and Confers Leupeptin Resistance
    Article Snippet: Paragraph title: Site-directed mutagenesis and DNA transfection. ... Whole-plasmid PCR was performed with PfuUltra Hotstart DNA polymerase (Stratagene); following DpnI digestion, the product was used to transform chemically competent TOP10 Escherichia coli (Invitrogen).

    Polymerase Chain Reaction:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: A CLAG3 Mutation in an Amphipathic Transmembrane Domain Alters Malaria Parasite Nutrient Channels and Confers Leupeptin Resistance
    Article Snippet: .. Whole-plasmid PCR was performed with PfuUltra Hotstart DNA polymerase (Stratagene); following DpnI digestion, the product was used to transform chemically competent TOP10 Escherichia coli (Invitrogen). ..

    Article Title: Human Bocavirus Can Be Cultured in Differentiated Human Airway Epithelial Cells ▿
    Article Snippet: .. Purified PCR products were cloned into the pCR2.1-TOPO TA vector (Invitrogen) and transformed in chemically competent TOP10 Escherichia coli (Invitrogen), according to manufacturer protocol. .. Transformants were directly analyzed via colony PCR with T7 and M13RP primers.

    Article Title: Efficient Site-Directed Saturation Mutagenesis Using Degenerate Oligonucleotides
    Article Snippet: However, we have not found it necessary to use partial overlapping primers, purified mutagenesis primers, or purified PCR reactions as they did. .. Reactions were used to transform 50 μL of chemically competent TOP10 E. coli (Invitrogen, Carlsbad, CA).

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: .. PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA). .. A staphylococcal origin of replication was introduced by cloning plasmid pRN5543 into the unique XbaI site on PCR 2.1-TOPO, and the constructs were moved into S. aureus RN4220 by electroporation ( ).

    DNA Sequencing:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Ligase Independent Cloning:

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: .. PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA). .. A staphylococcal origin of replication was introduced by cloning plasmid pRN5543 into the unique XbaI site on PCR 2.1-TOPO, and the constructs were moved into S. aureus RN4220 by electroporation ( ).

    Sequencing:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: Paragraph title: Cloning and sequencing of A . franciscana type 2 Hsp40 cDNA ... The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada).

    Article Title: Functional Aspects of Intrahepatic Hepatitis B Virus-specific T Cells Induced by Therapeutic DNA Vaccination
    Article Snippet: Plasmids were grown in competent TOP10 Escherichia coli (Life Technologies, Carlsbad, CA) and purified using Qiagen EndoFree Plasmid purification Kit according to the manufacturer's instructions (Qiagen, Hilden, Germany). .. All DNA plasmids were sequenced to ensure correct nucleotide sequence (Eurofins MWG Operon, Ebersberg, Germany).

    Recombinant:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Methylation:

    Article Title: Efficient Site-Directed Saturation Mutagenesis Using Degenerate Oligonucleotides
    Article Snippet: Reactions were cooled on ice and digested with 5 units of Dpn I for 1 h at 37ºC to cleave methylated and hemimethylated parental DNA, but not the newly synthesized mutant DNA molecules. .. Reactions were used to transform 50 μL of chemically competent TOP10 E. coli (Invitrogen, Carlsbad, CA).

    Mutagenesis:

    Article Title: A CLAG3 Mutation in an Amphipathic Transmembrane Domain Alters Malaria Parasite Nutrient Channels and Confers Leupeptin Resistance
    Article Snippet: Paragraph title: Site-directed mutagenesis and DNA transfection. ... Whole-plasmid PCR was performed with PfuUltra Hotstart DNA polymerase (Stratagene); following DpnI digestion, the product was used to transform chemically competent TOP10 Escherichia coli (Invitrogen).

    Article Title: Efficient Site-Directed Saturation Mutagenesis Using Degenerate Oligonucleotides
    Article Snippet: However, we have not found it necessary to use partial overlapping primers, purified mutagenesis primers, or purified PCR reactions as they did. .. Reactions were used to transform 50 μL of chemically competent TOP10 E. coli (Invitrogen, Carlsbad, CA).

    Isolation:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Size-exclusion Chromatography:

    Article Title: Efficient Site-Directed Saturation Mutagenesis Using Degenerate Oligonucleotides
    Article Snippet: Reactions were thermal cycled: 95ºC for 2 min, followed by 16 cycles of 95ºC for 30 sec, 55ºC for 1 min, and 68ºC for 10 min, then a final incubation of 68ºC for 10 min. .. Reactions were used to transform 50 μL of chemically competent TOP10 E. coli (Invitrogen, Carlsbad, CA).

    Purification:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: Human Bocavirus Can Be Cultured in Differentiated Human Airway Epithelial Cells ▿
    Article Snippet: .. Purified PCR products were cloned into the pCR2.1-TOPO TA vector (Invitrogen) and transformed in chemically competent TOP10 Escherichia coli (Invitrogen), according to manufacturer protocol. .. Transformants were directly analyzed via colony PCR with T7 and M13RP primers.

    Article Title: Efficient Site-Directed Saturation Mutagenesis Using Degenerate Oligonucleotides
    Article Snippet: However, we have not found it necessary to use partial overlapping primers, purified mutagenesis primers, or purified PCR reactions as they did. .. Reactions were used to transform 50 μL of chemically competent TOP10 E. coli (Invitrogen, Carlsbad, CA).

    Article Title: Functional Aspects of Intrahepatic Hepatitis B Virus-specific T Cells Induced by Therapeutic DNA Vaccination
    Article Snippet: .. Plasmids were grown in competent TOP10 Escherichia coli (Life Technologies, Carlsbad, CA) and purified using Qiagen EndoFree Plasmid purification Kit according to the manufacturer's instructions (Qiagen, Hilden, Germany). .. Plasmid DNA concentration was determined spectrophotometrically and the purified DNA was dissolved in sterile phosphate-buffered saline at concentrations of 2 mg/ml.

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: .. PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA). .. A staphylococcal origin of replication was introduced by cloning plasmid pRN5543 into the unique XbaI site on PCR 2.1-TOPO, and the constructs were moved into S. aureus RN4220 by electroporation ( ).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: RNA was quantified by measuring the absorbance at 260 nm and 0.1 μg mRNA was used as template for synthesizing cDNA with SuperScript® III First-Strand Synthesis Kit for RT-PCR (Invitrogen, Burlington, ON, Canada) and oligo dT20 following the manufacturer’s instructions. .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada).

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: RNA was quantified by measuring the absorbance at 260 nm and 0.1 μg mRNA was used as template for synthesizing cDNA with SuperScript® III First-Strand Synthesis Kit for RT-PCR (Invitrogen, Burlington, ON, Canada) and oligo dT20 following the manufacturer’s instructions. .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada).

    IA:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: Primers containing the restriction enzyme sites, BamH1 and HindIII, were designed based on sequence comparisons (Integrated DNA Technologies, Coralville, IA, USA) ( ) and used for the amplification of full length A . franciscana type 2 Hsp40 cDNA from 5 h cysts. .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada).

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: Primers containing the restriction enzyme sites, BamH1 and HindIII, were designed based on sequence comparisons (Integrated DNA Technologies, Coralville, IA, USA) ( ) and used for the amplification of full length A . franciscana type 2 Hsp40 cDNA from 5 h cysts. .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada).

    Plasmid Preparation:

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: Post-diapause synthesis of ArHsp40-2, a type 2 J-domain protein from Artemia franciscana, is developmentally regulated and induced by stress
    Article Snippet: .. The PCR products, resolved in 1.0% agarose gels and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega, Madison, WI, USA), was then ligated into a TA vector (TOPO TA Cloning Kit, Life Technologies, Burlington, ON, Canada) and used to transform competent TOP10 Escherichia coli (Invitrogen, Burlington, ON, Canada). .. Recombinant TA vectors containing cDNA inserts of the appropriate size were isolated with the GenElute™ Plasmid Miniprep Kit (Sigma-Aldrich, Oakville, ON, Canada), and the inserts were sequenced (DNA Sequencing Facility, Centre for Applied Genomics, Hospital for Sick Children, Toronto, ON, Canada) revealing a full-length type 2 Hsp40 cDNA termed ArHsp40-2 .

    Article Title: A CLAG3 Mutation in an Amphipathic Transmembrane Domain Alters Malaria Parasite Nutrient Channels and Confers Leupeptin Resistance
    Article Snippet: Previously described plasmid pHD22Y-120w-flag-PG1 ( ) was used as the template for site-directed mutagenesis with complementary primers (5′-ATGGTTTCATGTAT A CTTTTTGTTTTTTTGC-3′ and 5′-GCAAAAAAACAAAAAG T ATACATGAAACCAT-3′). .. Whole-plasmid PCR was performed with PfuUltra Hotstart DNA polymerase (Stratagene); following DpnI digestion, the product was used to transform chemically competent TOP10 Escherichia coli (Invitrogen).

    Article Title: Human Bocavirus Can Be Cultured in Differentiated Human Airway Epithelial Cells ▿
    Article Snippet: .. Purified PCR products were cloned into the pCR2.1-TOPO TA vector (Invitrogen) and transformed in chemically competent TOP10 Escherichia coli (Invitrogen), according to manufacturer protocol. .. Transformants were directly analyzed via colony PCR with T7 and M13RP primers.

    Article Title: Functional Aspects of Intrahepatic Hepatitis B Virus-specific T Cells Induced by Therapeutic DNA Vaccination
    Article Snippet: .. Plasmids were grown in competent TOP10 Escherichia coli (Life Technologies, Carlsbad, CA) and purified using Qiagen EndoFree Plasmid purification Kit according to the manufacturer's instructions (Qiagen, Hilden, Germany). .. Plasmid DNA concentration was determined spectrophotometrically and the purified DNA was dissolved in sterile phosphate-buffered saline at concentrations of 2 mg/ml.

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: .. PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA). .. A staphylococcal origin of replication was introduced by cloning plasmid pRN5543 into the unique XbaI site on PCR 2.1-TOPO, and the constructs were moved into S. aureus RN4220 by electroporation ( ).

    Concentration Assay:

    Article Title: Functional Aspects of Intrahepatic Hepatitis B Virus-specific T Cells Induced by Therapeutic DNA Vaccination
    Article Snippet: Plasmids were grown in competent TOP10 Escherichia coli (Life Technologies, Carlsbad, CA) and purified using Qiagen EndoFree Plasmid purification Kit according to the manufacturer's instructions (Qiagen, Hilden, Germany). .. Plasmid DNA concentration was determined spectrophotometrically and the purified DNA was dissolved in sterile phosphate-buffered saline at concentrations of 2 mg/ml.

    Gel Extraction:

    Article Title: A Subset of Staphylococcus aureus Strains Harboring Staphylococcal Cassette Chromosome mec (SCCmec) Type IV Is Deficient in CcrAB-Mediated SCCmec Excision †
    Article Snippet: .. PCR products were purified using the QIAquick gel extraction kit, ligated into the ligase-independent cloning site of the PCR 2.1-TOPO vector (Invitrogen, Carlsbad, CA), and transformed into chemically competent Top10 Escherichia coli (Invitrogen, Carlsbad, CA). .. A staphylococcal origin of replication was introduced by cloning plasmid pRN5543 into the unique XbaI site on PCR 2.1-TOPO, and the constructs were moved into S. aureus RN4220 by electroporation ( ).

    Homologous Recombination:

    Article Title: A CLAG3 Mutation in an Amphipathic Transmembrane Domain Alters Malaria Parasite Nutrient Channels and Confers Leupeptin Resistance
    Article Snippet: Whole-plasmid PCR was performed with PfuUltra Hotstart DNA polymerase (Stratagene); following DpnI digestion, the product was used to transform chemically competent TOP10 Escherichia coli (Invitrogen). .. The transfected culture was selected with 2.5 nM WR99210 and screened for homologous recombination by PCR.

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    Thermo Fisher one shot top10 chemically competent e coli
    One Shot Top10 Chemically Competent E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    one shot top10 chemically competent e coli - by Bioz Stars, 2020-01
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    MultiShot StripWell TOP10 chemically competent E coli cells are cloning competent cells that packaged in a rack containing 12 strips of 8 tubes to increase productivity for medium throughput bacterial
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