bp clonase ii enzyme mix Thermo Fisher Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher gateway bp clonase ii enzyme mix
    Gateway® BP and LR reactions. BP and LR <t>Clonase®</t> facilitate the recombination between att B and att P and between att L and att R, generating att L and att B sequences, respectively.
    Gateway Bp Clonase Ii Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1960 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gateway bp clonase ii enzyme mix/product/Thermo Fisher
    Average 99 stars, based on 1960 article reviews
    Price from $9.99 to $1999.99
    gateway bp clonase ii enzyme mix - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher bp clonase ii enzyme mix
    Drosophila destination vector and two-fragment schematic for Gateway MultiSite cloning. A) The Drosophila destination vector pDESThaw. This destination vector contains an hsp70 polyadenylation sequence, a PhiC31 attB site for PhiC31 integrase-mediate site-specific transgenesis, and mini- white as a transformation marker. Appropriate combinations of two, three, or four entry clones recombine in a position and orientation-specific manner with ampicillin-resistant pDESThaw in the LR reaction to generate expression clones. The LR reactions for two, three, and four fragment Gateway MultiSite recombination all use the LR <t>Clonase</t> II Plus enzyme mix. Both the BP and LR reactions take advantage of the Gateway cassette that includes a chloramphenicol resistance marker and the ccdB gene. The Gateway cassette is located between the attP sites in the pDONR vectors ( Figure S1 ) and between the attR1 and attR2 sites of the pDESThaw destination vector. The ccdB gene is toxic to any bacterial strain that does not contain a genetic suppressor including most common laboratory bacterial strains used for cloning such as DH10B and DH5α . ccdB containing clones were propagated using the now discontinued bacterial strain DB3.1 that has been replaced by Invitrogen with strain ccdB Survival 2 T1R. When BP and LR reactions are transformed into non-ccdB suppressor strains, only bacteria containing clones in which the Gateway cassette has been recombined out (and presumably the fragment(s) of interest recombined in) survive on kanamycin or ampicillin-selective plates. This results in a low frequency of colonies that do not contain the insert(s) of interest. B) Schematic diagram of two-fragment Gateway MultiSite recombination cloning. Fragment 1 and fragment 2 are amplified by PCR using oligonucleotides that incorporate flanking attB1 and attB5r sites in fragment 1 and flanking attB5 and attB2 sites in fragment 2. Fragment 1 is combined with pDONR 221 P1-P5r and fragment 2 is combined with pDONR 221 P5-P2 in separate BP reactions. The products of the BP reactions are pENTR attL1-Frag1-attR5 and pENTR attL5-Frag2-attL2. In the LR reaction both of these entry clones are combined with a destination vector to produce an expression clone containing fragment 1 and fragment 2 in a position and orientation-specific manner. Note that pENTR L1-R5 entry clones are also used in four-fragment Gateway MultiSite cloning. Schematic modified from the Invitrogen MultiSite Gateway Pro user manual.
    Bp Clonase Ii Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1770 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bp clonase ii enzyme mix/product/Thermo Fisher
    Average 99 stars, based on 1770 article reviews
    Price from $9.99 to $1999.99
    bp clonase ii enzyme mix - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher lr clonase ii enzyme mix
    Drosophila destination vector and two-fragment schematic for Gateway MultiSite cloning. A) The Drosophila destination vector pDESThaw. This destination vector contains an hsp70 polyadenylation sequence, a PhiC31 attB site for PhiC31 integrase-mediate site-specific transgenesis, and mini- white as a transformation marker. Appropriate combinations of two, three, or four entry clones recombine in a position and orientation-specific manner with ampicillin-resistant pDESThaw in the LR reaction to generate expression clones. The LR reactions for two, three, and four fragment Gateway MultiSite recombination all use the LR <t>Clonase</t> II Plus enzyme mix. Both the BP and LR reactions take advantage of the Gateway cassette that includes a chloramphenicol resistance marker and the ccdB gene. The Gateway cassette is located between the attP sites in the pDONR vectors ( Figure S1 ) and between the attR1 and attR2 sites of the pDESThaw destination vector. The ccdB gene is toxic to any bacterial strain that does not contain a genetic suppressor including most common laboratory bacterial strains used for cloning such as DH10B and DH5α . ccdB containing clones were propagated using the now discontinued bacterial strain DB3.1 that has been replaced by Invitrogen with strain ccdB Survival 2 T1R. When BP and LR reactions are transformed into non-ccdB suppressor strains, only bacteria containing clones in which the Gateway cassette has been recombined out (and presumably the fragment(s) of interest recombined in) survive on kanamycin or ampicillin-selective plates. This results in a low frequency of colonies that do not contain the insert(s) of interest. B) Schematic diagram of two-fragment Gateway MultiSite recombination cloning. Fragment 1 and fragment 2 are amplified by PCR using oligonucleotides that incorporate flanking attB1 and attB5r sites in fragment 1 and flanking attB5 and attB2 sites in fragment 2. Fragment 1 is combined with pDONR 221 P1-P5r and fragment 2 is combined with pDONR 221 P5-P2 in separate BP reactions. The products of the BP reactions are pENTR attL1-Frag1-attR5 and pENTR attL5-Frag2-attL2. In the LR reaction both of these entry clones are combined with a destination vector to produce an expression clone containing fragment 1 and fragment 2 in a position and orientation-specific manner. Note that pENTR L1-R5 entry clones are also used in four-fragment Gateway MultiSite cloning. Schematic modified from the Invitrogen MultiSite Gateway Pro user manual.
    Lr Clonase Ii Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2033 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lr clonase ii enzyme mix/product/Thermo Fisher
    Average 99 stars, based on 2033 article reviews
    Price from $9.99 to $1999.99
    lr clonase ii enzyme mix - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    91
    Thermo Fisher camv 35s promoter
    Drosophila destination vector and two-fragment schematic for Gateway MultiSite cloning. A) The Drosophila destination vector pDESThaw. This destination vector contains an hsp70 polyadenylation sequence, a PhiC31 attB site for PhiC31 integrase-mediate site-specific transgenesis, and mini- white as a transformation marker. Appropriate combinations of two, three, or four entry clones recombine in a position and orientation-specific manner with ampicillin-resistant pDESThaw in the LR reaction to generate expression clones. The LR reactions for two, three, and four fragment Gateway MultiSite recombination all use the LR <t>Clonase</t> II Plus enzyme mix. Both the BP and LR reactions take advantage of the Gateway cassette that includes a chloramphenicol resistance marker and the ccdB gene. The Gateway cassette is located between the attP sites in the pDONR vectors ( Figure S1 ) and between the attR1 and attR2 sites of the pDESThaw destination vector. The ccdB gene is toxic to any bacterial strain that does not contain a genetic suppressor including most common laboratory bacterial strains used for cloning such as DH10B and DH5α . ccdB containing clones were propagated using the now discontinued bacterial strain DB3.1 that has been replaced by Invitrogen with strain ccdB Survival 2 T1R. When BP and LR reactions are transformed into non-ccdB suppressor strains, only bacteria containing clones in which the Gateway cassette has been recombined out (and presumably the fragment(s) of interest recombined in) survive on kanamycin or ampicillin-selective plates. This results in a low frequency of colonies that do not contain the insert(s) of interest. B) Schematic diagram of two-fragment Gateway MultiSite recombination cloning. Fragment 1 and fragment 2 are amplified by PCR using oligonucleotides that incorporate flanking attB1 and attB5r sites in fragment 1 and flanking attB5 and attB2 sites in fragment 2. Fragment 1 is combined with pDONR 221 P1-P5r and fragment 2 is combined with pDONR 221 P5-P2 in separate BP reactions. The products of the BP reactions are pENTR attL1-Frag1-attR5 and pENTR attL5-Frag2-attL2. In the LR reaction both of these entry clones are combined with a destination vector to produce an expression clone containing fragment 1 and fragment 2 in a position and orientation-specific manner. Note that pENTR L1-R5 entry clones are also used in four-fragment Gateway MultiSite cloning. Schematic modified from the Invitrogen MultiSite Gateway Pro user manual.
    Camv 35s Promoter, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 426 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/camv 35s promoter/product/Thermo Fisher
    Average 91 stars, based on 426 article reviews
    Price from $9.99 to $1999.99
    camv 35s promoter - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    Image Search Results


    Gateway® BP and LR reactions. BP and LR Clonase® facilitate the recombination between att B and att P and between att L and att R, generating att L and att B sequences, respectively.

    Journal: Current protocols in molecular biology / edited by Frederick M. Ausubel ... [et al.]

    Article Title: Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes

    doi: 10.1002/0471142727.mb0320s110

    Figure Lengend Snippet: Gateway® BP and LR reactions. BP and LR Clonase® facilitate the recombination between att B and att P and between att L and att R, generating att L and att B sequences, respectively.

    Article Snippet: 150 ng/µl entry vector (e.g., pDONR™221; Life Technologies) Gateway® BP Clonase® II enzyme mix (Life Technologies) 10 to 50 ng/µl att B-PCR product prepared according to the Gateway® scheme (Basic Protocol 1) PCR-grade H2 O LB plates supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) LB medium supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) 50% (v/v) glycerol, sterile Plasmid DNA isolation kit (e.g., Qiagen; optional) 96-well plate or 1.5 ml microcentrifuge tubes Multichannel pipettor 25°C incubator Additional reagents and equipment for growth of T1 phage resistant (T1R ) E. coli competent cells (e.g. One Shot® MAX Efficiency® DH5α™ competent cells; Life Technologies) ( unit 1.8 ), DNA miniprep ( unit 1.6 or commercial DNA prep kit), and DNA sequencing (Chapter 7).

    Techniques:

    Drosophila destination vector and two-fragment schematic for Gateway MultiSite cloning. A) The Drosophila destination vector pDESThaw. This destination vector contains an hsp70 polyadenylation sequence, a PhiC31 attB site for PhiC31 integrase-mediate site-specific transgenesis, and mini- white as a transformation marker. Appropriate combinations of two, three, or four entry clones recombine in a position and orientation-specific manner with ampicillin-resistant pDESThaw in the LR reaction to generate expression clones. The LR reactions for two, three, and four fragment Gateway MultiSite recombination all use the LR Clonase II Plus enzyme mix. Both the BP and LR reactions take advantage of the Gateway cassette that includes a chloramphenicol resistance marker and the ccdB gene. The Gateway cassette is located between the attP sites in the pDONR vectors ( Figure S1 ) and between the attR1 and attR2 sites of the pDESThaw destination vector. The ccdB gene is toxic to any bacterial strain that does not contain a genetic suppressor including most common laboratory bacterial strains used for cloning such as DH10B and DH5α . ccdB containing clones were propagated using the now discontinued bacterial strain DB3.1 that has been replaced by Invitrogen with strain ccdB Survival 2 T1R. When BP and LR reactions are transformed into non-ccdB suppressor strains, only bacteria containing clones in which the Gateway cassette has been recombined out (and presumably the fragment(s) of interest recombined in) survive on kanamycin or ampicillin-selective plates. This results in a low frequency of colonies that do not contain the insert(s) of interest. B) Schematic diagram of two-fragment Gateway MultiSite recombination cloning. Fragment 1 and fragment 2 are amplified by PCR using oligonucleotides that incorporate flanking attB1 and attB5r sites in fragment 1 and flanking attB5 and attB2 sites in fragment 2. Fragment 1 is combined with pDONR 221 P1-P5r and fragment 2 is combined with pDONR 221 P5-P2 in separate BP reactions. The products of the BP reactions are pENTR attL1-Frag1-attR5 and pENTR attL5-Frag2-attL2. In the LR reaction both of these entry clones are combined with a destination vector to produce an expression clone containing fragment 1 and fragment 2 in a position and orientation-specific manner. Note that pENTR L1-R5 entry clones are also used in four-fragment Gateway MultiSite cloning. Schematic modified from the Invitrogen MultiSite Gateway Pro user manual.

    Journal: PLoS ONE

    Article Title: A Gateway MultiSite Recombination Cloning Toolkit

    doi: 10.1371/journal.pone.0024531

    Figure Lengend Snippet: Drosophila destination vector and two-fragment schematic for Gateway MultiSite cloning. A) The Drosophila destination vector pDESThaw. This destination vector contains an hsp70 polyadenylation sequence, a PhiC31 attB site for PhiC31 integrase-mediate site-specific transgenesis, and mini- white as a transformation marker. Appropriate combinations of two, three, or four entry clones recombine in a position and orientation-specific manner with ampicillin-resistant pDESThaw in the LR reaction to generate expression clones. The LR reactions for two, three, and four fragment Gateway MultiSite recombination all use the LR Clonase II Plus enzyme mix. Both the BP and LR reactions take advantage of the Gateway cassette that includes a chloramphenicol resistance marker and the ccdB gene. The Gateway cassette is located between the attP sites in the pDONR vectors ( Figure S1 ) and between the attR1 and attR2 sites of the pDESThaw destination vector. The ccdB gene is toxic to any bacterial strain that does not contain a genetic suppressor including most common laboratory bacterial strains used for cloning such as DH10B and DH5α . ccdB containing clones were propagated using the now discontinued bacterial strain DB3.1 that has been replaced by Invitrogen with strain ccdB Survival 2 T1R. When BP and LR reactions are transformed into non-ccdB suppressor strains, only bacteria containing clones in which the Gateway cassette has been recombined out (and presumably the fragment(s) of interest recombined in) survive on kanamycin or ampicillin-selective plates. This results in a low frequency of colonies that do not contain the insert(s) of interest. B) Schematic diagram of two-fragment Gateway MultiSite recombination cloning. Fragment 1 and fragment 2 are amplified by PCR using oligonucleotides that incorporate flanking attB1 and attB5r sites in fragment 1 and flanking attB5 and attB2 sites in fragment 2. Fragment 1 is combined with pDONR 221 P1-P5r and fragment 2 is combined with pDONR 221 P5-P2 in separate BP reactions. The products of the BP reactions are pENTR attL1-Frag1-attR5 and pENTR attL5-Frag2-attL2. In the LR reaction both of these entry clones are combined with a destination vector to produce an expression clone containing fragment 1 and fragment 2 in a position and orientation-specific manner. Note that pENTR L1-R5 entry clones are also used in four-fragment Gateway MultiSite cloning. Schematic modified from the Invitrogen MultiSite Gateway Pro user manual.

    Article Snippet: All BP reactions were performed using the BP Clonase II enzyme mix (Invitrogen-Cat # 11789-020) in 5 µl volumes (half the manufacturer's recommendation) for 1 hr to overnight at room temperature (Proteinase K digestion omitted), run through a spin column (Zymo Research Cat # D4104), eluted in 6 µl of water, electroporated (2 µl) into 50 µl DH10B electrocompetent cells prepared in our laboratory, and plated on LB kanamycin plates (50 µg/ml).

    Techniques: Plasmid Preparation, Clone Assay, Sequencing, Transformation Assay, Marker, Expressing, Amplification, Polymerase Chain Reaction, Modification