gateway polymerase chain reaction pcr cloning strategy  (Thermo Fisher)


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    Name:
    PCR Cloning System
    Description:
    Gateway Technology enables rapid cloning of one or more genes into virtually any protein expression system Once you have an entry clone you can recombine your gene of interest into a variety of expression vectors adapted for use in Gateway Technology The PCR Cloning System with Gateway Technology is designed to create an entry clone following PCR with attB modified custom primers Figure 1 The PCR product is directionally cloned with efficiencies typically 99 The PCR Cloning System with Gateway Technology offers • Rapid efficient directional cloning of PCR products• Versatility to use any amplification enzyme• Cloning without the need for restriction endonucleases or ligase• Selection of entry clones with kanamycin or Zeocin In addition the pDONR 221 and pDONR Zeo vectors have a pUC origin for high plasmid yields and universal M13 sequencing sites for ease of use
    Catalog Number:
    12535029
    Price:
    None
    Applications:
    Cloning|Donor Vectors|Gateway Cloning
    Category:
    DNA Vectors Clones Purified Nucleic Acids Libraries
    Buy from Supplier


    Structured Review

    Thermo Fisher gateway polymerase chain reaction pcr cloning strategy
    Gateway Technology enables rapid cloning of one or more genes into virtually any protein expression system Once you have an entry clone you can recombine your gene of interest into a variety of expression vectors adapted for use in Gateway Technology The PCR Cloning System with Gateway Technology is designed to create an entry clone following PCR with attB modified custom primers Figure 1 The PCR product is directionally cloned with efficiencies typically 99 The PCR Cloning System with Gateway Technology offers • Rapid efficient directional cloning of PCR products• Versatility to use any amplification enzyme• Cloning without the need for restriction endonucleases or ligase• Selection of entry clones with kanamycin or Zeocin In addition the pDONR 221 and pDONR Zeo vectors have a pUC origin for high plasmid yields and universal M13 sequencing sites for ease of use
    https://www.bioz.com/result/gateway polymerase chain reaction pcr cloning strategy/product/Thermo Fisher
    Average 94 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    gateway polymerase chain reaction pcr cloning strategy - by Bioz Stars, 2020-07
    94/100 stars

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    Related Articles

    Clone Assay:

    Article Title: New insight into HCV E1/E2 region of genotype 4a
    Article Snippet: .. Cloning Purified PCR products were directly ligated into pCR2.1- TOPO plasmid (Invitrogen, Carlsbad, CA, USA) and then chemically transformed into One Shot Top10 Escherichia coli (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s instructions. ..

    Article Title: Genetic characterization of the wboA gene from the predominant biovars of Brucella isolates in Iran
    Article Snippet: .. Cloning and sequencing PCR products were cloned using an Ins TA clone PCR Cloning Kit (Thermo Scientific) according to the manufacturer’s instructions. .. The transformation was performed using competent E. coli DH5 α cells.

    Article Title: A time-resolved immunoassay to measure serum antibodies to the rotavirus VP6 capsid protein
    Article Snippet: .. The rVP6 DNAs were cloned into a shuttle vector (pDONR221) using the primers outlined in and transferred into the baculovirus expression vector (pDEST17), using the Gateway® PCR cloning system (Invitrogen). ..

    Article Title: Vitamin B1 diversity and characterization of biosynthesis genes in cassava
    Article Snippet: .. PCR products were run on a 1% (w/v) agarose gel and the different bands, corresponding to THIC 3ʹ-UTR splice variants, were extracted, ligated into the pJET1.2/blunt vector with the Clone PCR cloning Kit (Thermo Scientific) according to the manufacturer’s instructions, and transformed into thermocompetent Escherichia coli cells. .. Two to six clones were sequenced for each 3ʹ-UTR splicing variant using the Sanger sequencing method.

    Article Title: Development of Antigen-Specific Memory CD8+ T Cells Following Live-Attenuated Chimeric West Nile Virus Vaccination
    Article Snippet: .. The WNV E sequence was amplified from the full-length WNV complementary DNA [ ] by use of GeneAmp XL polymerase chain reaction (PCR; Applied Biosystems) and cloned into the donor vector pDONR221 by use of a PCR cloning system with Gateway Technology (Invitrogen). .. The vaccinia transfer plasmid pMJ601 was modified to facilitate insertion of the WNV E gene (Gateway Vector conversion system; Invitrogen).

    Article Title: Knocking-Down Meloidogyne incognita Proteases by Plant-Delivered dsRNA Has Negative Pleiotropic Effect on Nematode Vigor
    Article Snippet: .. All fragments and fusion were subcloned into the pDONR vector™ 221, using BP clonase (PCR Cloning System with Gateway® Technology Kit, Invitrogen, USA), and then transferred again by recombination, using LR clonase enzyme, for the binary vector pK7GWIWG2 (I) [ ] used for expressing dsRNA in plants. .. Generation and selection of transgenic RNAi tobacco lines dsRNA constructs were inserted into A. tumefaciens strain GV3101 using a standard electroporation method and plated on LB medium containing rifampicin (100 µg ml-1 ), kanamycin (50 µg ml-1 ) and streptomycin (300 µg ml-1 ).

    Amplification:

    Article Title: Development of Antigen-Specific Memory CD8+ T Cells Following Live-Attenuated Chimeric West Nile Virus Vaccination
    Article Snippet: .. The WNV E sequence was amplified from the full-length WNV complementary DNA [ ] by use of GeneAmp XL polymerase chain reaction (PCR; Applied Biosystems) and cloned into the donor vector pDONR221 by use of a PCR cloning system with Gateway Technology (Invitrogen). .. The vaccinia transfer plasmid pMJ601 was modified to facilitate insertion of the WNV E gene (Gateway Vector conversion system; Invitrogen).

    Agarose Gel Electrophoresis:

    Article Title: Vitamin B1 diversity and characterization of biosynthesis genes in cassava
    Article Snippet: .. PCR products were run on a 1% (w/v) agarose gel and the different bands, corresponding to THIC 3ʹ-UTR splice variants, were extracted, ligated into the pJET1.2/blunt vector with the Clone PCR cloning Kit (Thermo Scientific) according to the manufacturer’s instructions, and transformed into thermocompetent Escherichia coli cells. .. Two to six clones were sequenced for each 3ʹ-UTR splicing variant using the Sanger sequencing method.

    Construct:

    Article Title: Recombinant protein rVP1 upregulates BECN1-independent autophagy, MAPK1/3 phosphorylation and MMP9 activity via WIPI1/WIPI2 to promote macrophage migration
    Article Snippet: .. The pDEST- DsRed-Wipi1 and pDEST- DsRed-WIPI1 plasmid was constructed using a Gateway recombination system (Invitrogen, 12535-029) according to the manufacturer’s instructions. .. Briefly, Wipi1 or WIPI1 cDNA was amplified by PCR from the pCMV-SPORT6- Wipi1 or pCMV-SPORT6- WIPI1 plasmid (Open Biosystems, clone ID 5322999 for Wipi1 and ID6055425 for WIPI1 ) respectively and subcloned into the Gateway pDONR vector to generate the pDONR- Wipi1 and pDONR- WIPI1 entry clone.

    Purification:

    Article Title: New insight into HCV E1/E2 region of genotype 4a
    Article Snippet: .. Cloning Purified PCR products were directly ligated into pCR2.1- TOPO plasmid (Invitrogen, Carlsbad, CA, USA) and then chemically transformed into One Shot Top10 Escherichia coli (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s instructions. ..

    Polymerase Chain Reaction:

    Article Title: New insight into HCV E1/E2 region of genotype 4a
    Article Snippet: .. Cloning Purified PCR products were directly ligated into pCR2.1- TOPO plasmid (Invitrogen, Carlsbad, CA, USA) and then chemically transformed into One Shot Top10 Escherichia coli (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s instructions. ..

    Article Title: Genetic characterization of the wboA gene from the predominant biovars of Brucella isolates in Iran
    Article Snippet: .. Cloning and sequencing PCR products were cloned using an Ins TA clone PCR Cloning Kit (Thermo Scientific) according to the manufacturer’s instructions. .. The transformation was performed using competent E. coli DH5 α cells.

    Article Title: A time-resolved immunoassay to measure serum antibodies to the rotavirus VP6 capsid protein
    Article Snippet: .. The rVP6 DNAs were cloned into a shuttle vector (pDONR221) using the primers outlined in and transferred into the baculovirus expression vector (pDEST17), using the Gateway® PCR cloning system (Invitrogen). ..

    Article Title: Vitamin B1 diversity and characterization of biosynthesis genes in cassava
    Article Snippet: .. PCR products were run on a 1% (w/v) agarose gel and the different bands, corresponding to THIC 3ʹ-UTR splice variants, were extracted, ligated into the pJET1.2/blunt vector with the Clone PCR cloning Kit (Thermo Scientific) according to the manufacturer’s instructions, and transformed into thermocompetent Escherichia coli cells. .. Two to six clones were sequenced for each 3ʹ-UTR splicing variant using the Sanger sequencing method.

    Article Title: Development of Antigen-Specific Memory CD8+ T Cells Following Live-Attenuated Chimeric West Nile Virus Vaccination
    Article Snippet: .. The WNV E sequence was amplified from the full-length WNV complementary DNA [ ] by use of GeneAmp XL polymerase chain reaction (PCR; Applied Biosystems) and cloned into the donor vector pDONR221 by use of a PCR cloning system with Gateway Technology (Invitrogen). .. The vaccinia transfer plasmid pMJ601 was modified to facilitate insertion of the WNV E gene (Gateway Vector conversion system; Invitrogen).

    Article Title: Knocking-Down Meloidogyne incognita Proteases by Plant-Delivered dsRNA Has Negative Pleiotropic Effect on Nematode Vigor
    Article Snippet: .. All fragments and fusion were subcloned into the pDONR vector™ 221, using BP clonase (PCR Cloning System with Gateway® Technology Kit, Invitrogen, USA), and then transferred again by recombination, using LR clonase enzyme, for the binary vector pK7GWIWG2 (I) [ ] used for expressing dsRNA in plants. .. Generation and selection of transgenic RNAi tobacco lines dsRNA constructs were inserted into A. tumefaciens strain GV3101 using a standard electroporation method and plated on LB medium containing rifampicin (100 µg ml-1 ), kanamycin (50 µg ml-1 ) and streptomycin (300 µg ml-1 ).

    Expressing:

    Article Title: A time-resolved immunoassay to measure serum antibodies to the rotavirus VP6 capsid protein
    Article Snippet: .. The rVP6 DNAs were cloned into a shuttle vector (pDONR221) using the primers outlined in and transferred into the baculovirus expression vector (pDEST17), using the Gateway® PCR cloning system (Invitrogen). ..

    Article Title: Knocking-Down Meloidogyne incognita Proteases by Plant-Delivered dsRNA Has Negative Pleiotropic Effect on Nematode Vigor
    Article Snippet: .. All fragments and fusion were subcloned into the pDONR vector™ 221, using BP clonase (PCR Cloning System with Gateway® Technology Kit, Invitrogen, USA), and then transferred again by recombination, using LR clonase enzyme, for the binary vector pK7GWIWG2 (I) [ ] used for expressing dsRNA in plants. .. Generation and selection of transgenic RNAi tobacco lines dsRNA constructs were inserted into A. tumefaciens strain GV3101 using a standard electroporation method and plated on LB medium containing rifampicin (100 µg ml-1 ), kanamycin (50 µg ml-1 ) and streptomycin (300 µg ml-1 ).

    Sequencing:

    Article Title: Genetic characterization of the wboA gene from the predominant biovars of Brucella isolates in Iran
    Article Snippet: .. Cloning and sequencing PCR products were cloned using an Ins TA clone PCR Cloning Kit (Thermo Scientific) according to the manufacturer’s instructions. .. The transformation was performed using competent E. coli DH5 α cells.

    Article Title: Development of Antigen-Specific Memory CD8+ T Cells Following Live-Attenuated Chimeric West Nile Virus Vaccination
    Article Snippet: .. The WNV E sequence was amplified from the full-length WNV complementary DNA [ ] by use of GeneAmp XL polymerase chain reaction (PCR; Applied Biosystems) and cloned into the donor vector pDONR221 by use of a PCR cloning system with Gateway Technology (Invitrogen). .. The vaccinia transfer plasmid pMJ601 was modified to facilitate insertion of the WNV E gene (Gateway Vector conversion system; Invitrogen).

    Transformation Assay:

    Article Title: New insight into HCV E1/E2 region of genotype 4a
    Article Snippet: .. Cloning Purified PCR products were directly ligated into pCR2.1- TOPO plasmid (Invitrogen, Carlsbad, CA, USA) and then chemically transformed into One Shot Top10 Escherichia coli (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s instructions. ..

    Article Title: Vitamin B1 diversity and characterization of biosynthesis genes in cassava
    Article Snippet: .. PCR products were run on a 1% (w/v) agarose gel and the different bands, corresponding to THIC 3ʹ-UTR splice variants, were extracted, ligated into the pJET1.2/blunt vector with the Clone PCR cloning Kit (Thermo Scientific) according to the manufacturer’s instructions, and transformed into thermocompetent Escherichia coli cells. .. Two to six clones were sequenced for each 3ʹ-UTR splicing variant using the Sanger sequencing method.

    Plasmid Preparation:

    Article Title: New insight into HCV E1/E2 region of genotype 4a
    Article Snippet: .. Cloning Purified PCR products were directly ligated into pCR2.1- TOPO plasmid (Invitrogen, Carlsbad, CA, USA) and then chemically transformed into One Shot Top10 Escherichia coli (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s instructions. ..

    Article Title: Recombinant protein rVP1 upregulates BECN1-independent autophagy, MAPK1/3 phosphorylation and MMP9 activity via WIPI1/WIPI2 to promote macrophage migration
    Article Snippet: .. The pDEST- DsRed-Wipi1 and pDEST- DsRed-WIPI1 plasmid was constructed using a Gateway recombination system (Invitrogen, 12535-029) according to the manufacturer’s instructions. .. Briefly, Wipi1 or WIPI1 cDNA was amplified by PCR from the pCMV-SPORT6- Wipi1 or pCMV-SPORT6- WIPI1 plasmid (Open Biosystems, clone ID 5322999 for Wipi1 and ID6055425 for WIPI1 ) respectively and subcloned into the Gateway pDONR vector to generate the pDONR- Wipi1 and pDONR- WIPI1 entry clone.

    Article Title: A time-resolved immunoassay to measure serum antibodies to the rotavirus VP6 capsid protein
    Article Snippet: .. The rVP6 DNAs were cloned into a shuttle vector (pDONR221) using the primers outlined in and transferred into the baculovirus expression vector (pDEST17), using the Gateway® PCR cloning system (Invitrogen). ..

    Article Title: Vitamin B1 diversity and characterization of biosynthesis genes in cassava
    Article Snippet: .. PCR products were run on a 1% (w/v) agarose gel and the different bands, corresponding to THIC 3ʹ-UTR splice variants, were extracted, ligated into the pJET1.2/blunt vector with the Clone PCR cloning Kit (Thermo Scientific) according to the manufacturer’s instructions, and transformed into thermocompetent Escherichia coli cells. .. Two to six clones were sequenced for each 3ʹ-UTR splicing variant using the Sanger sequencing method.

    Article Title: Development of Antigen-Specific Memory CD8+ T Cells Following Live-Attenuated Chimeric West Nile Virus Vaccination
    Article Snippet: .. The WNV E sequence was amplified from the full-length WNV complementary DNA [ ] by use of GeneAmp XL polymerase chain reaction (PCR; Applied Biosystems) and cloned into the donor vector pDONR221 by use of a PCR cloning system with Gateway Technology (Invitrogen). .. The vaccinia transfer plasmid pMJ601 was modified to facilitate insertion of the WNV E gene (Gateway Vector conversion system; Invitrogen).

    Article Title: Knocking-Down Meloidogyne incognita Proteases by Plant-Delivered dsRNA Has Negative Pleiotropic Effect on Nematode Vigor
    Article Snippet: .. All fragments and fusion were subcloned into the pDONR vector™ 221, using BP clonase (PCR Cloning System with Gateway® Technology Kit, Invitrogen, USA), and then transferred again by recombination, using LR clonase enzyme, for the binary vector pK7GWIWG2 (I) [ ] used for expressing dsRNA in plants. .. Generation and selection of transgenic RNAi tobacco lines dsRNA constructs were inserted into A. tumefaciens strain GV3101 using a standard electroporation method and plated on LB medium containing rifampicin (100 µg ml-1 ), kanamycin (50 µg ml-1 ) and streptomycin (300 µg ml-1 ).

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