max efficiency dh5α competent cells  (Thermo Fisher)


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    Name:
    MAX Efficiency DH5α Competent Cells
    Description:
    MAX Efficiency DH5α Competent Cells are a well known versatile strain that can be used in many everyday cloning applications In addition to supporting blue white screening recA1 and endA1 mutations in DH5α cells increase insert stability and improve the quality of plasmid DNA prepared from minipreps Features of MAX Efficiency DH5α Competent Cells include • Transformation efficiency up to 1 × 109 transformants µg plasmid DNA• High plasmid yield from the DH5α endA1 E coli strain• Blue white screening capable lacZΔM15 • Greater insert stability recA1 About MAX Efficiency DH5α Competent CellsMAX Efficiency DH5α Competent Cells have been prepared by a proprietary modification of the procedure of Hanahan 1983 These cells are suitable for the construction of gene banks or for the generation of cDNA libraries using plasmid derived vectors The Φ80lacZΔM15 marker provides α complementation of the β galactosidase gene from pUC or similar vectors and can therefore be used for blue white screening of colonies on bacterial plates containing Bluo gal or X gal DH5α cells are capable of being transformed efficiently with large plasmids and can also serve as a host for the M13mp cloning vectors if a lawn of DH5α FT DH5αF DH5αF IQ JM101 or JM107 is provided to allow plaque formation Genotype F Φ80lacZΔM15 Δ lacZYA argF U169 recA1 endA1 hsdR17 rk mk phoA supE44 λ thi 1 gyrA96 relA1Find the strain and format that you needDH5α cells are available with a variety of transformation efficiencies and in both electrocompetent and chemically competent formats and in single use aliquots • Subcloning Efficiency DH5α Competent Cells our most economical competent cell for everyday use• Library Efficiency DH5α Competent Cells economical and for use with difficult to transform DNA• MAX Efficiency DH5α T1R Competent Cells T1 phage resistant 109 transformants µg plasmid DNA• One Shot MAX Efficiency DH5α T1R Competent Cells single use format T1 phage resistant 109 transformants µg plasmid DNA• ElectroMAX DH5α Competent Cells electrocompetent with 1010 transformants µg plasmid DNA and ideal for cDNA libraries or larger inserts
    Catalog Number:
    18258012
    Price:
    None
    Applications:
    Chemically Competent Cells for Cloning|Cloning|Transformation
    Category:
    Competent Cells Strains
    Buy from Supplier


    Structured Review

    Thermo Fisher max efficiency dh5α competent cells
    Diagnostic PCR confirms the deletion of TK0566 from the T. <t>kodakarensis</t> genome The same primer pairs are used on the <t>TS559</t> genome as the deletion genome. The size shift using primers A/B corresponds to the deletion of TK0566 while the absence of products for the reactions using C/D and E/F confirms the gene has been deleted from its native locus. The absence of products using primers C/D demonstrates that TK0566 is not present anywhere in the genome of the deletion strain.
    MAX Efficiency DH5α Competent Cells are a well known versatile strain that can be used in many everyday cloning applications In addition to supporting blue white screening recA1 and endA1 mutations in DH5α cells increase insert stability and improve the quality of plasmid DNA prepared from minipreps Features of MAX Efficiency DH5α Competent Cells include • Transformation efficiency up to 1 × 109 transformants µg plasmid DNA• High plasmid yield from the DH5α endA1 E coli strain• Blue white screening capable lacZΔM15 • Greater insert stability recA1 About MAX Efficiency DH5α Competent CellsMAX Efficiency DH5α Competent Cells have been prepared by a proprietary modification of the procedure of Hanahan 1983 These cells are suitable for the construction of gene banks or for the generation of cDNA libraries using plasmid derived vectors The Φ80lacZΔM15 marker provides α complementation of the β galactosidase gene from pUC or similar vectors and can therefore be used for blue white screening of colonies on bacterial plates containing Bluo gal or X gal DH5α cells are capable of being transformed efficiently with large plasmids and can also serve as a host for the M13mp cloning vectors if a lawn of DH5α FT DH5αF DH5αF IQ JM101 or JM107 is provided to allow plaque formation Genotype F Φ80lacZΔM15 Δ lacZYA argF U169 recA1 endA1 hsdR17 rk mk phoA supE44 λ thi 1 gyrA96 relA1Find the strain and format that you needDH5α cells are available with a variety of transformation efficiencies and in both electrocompetent and chemically competent formats and in single use aliquots • Subcloning Efficiency DH5α Competent Cells our most economical competent cell for everyday use• Library Efficiency DH5α Competent Cells economical and for use with difficult to transform DNA• MAX Efficiency DH5α T1R Competent Cells T1 phage resistant 109 transformants µg plasmid DNA• One Shot MAX Efficiency DH5α T1R Competent Cells single use format T1 phage resistant 109 transformants µg plasmid DNA• ElectroMAX DH5α Competent Cells electrocompetent with 1010 transformants µg plasmid DNA and ideal for cDNA libraries or larger inserts
    https://www.bioz.com/result/max efficiency dh5α competent cells/product/Thermo Fisher
    Average 96 stars, based on 65 article reviews
    Price from $9.99 to $1999.99
    max efficiency dh5α competent cells - by Bioz Stars, 2020-08
    96/100 stars

    Images

    1) Product Images from "Markerless Gene Editing in the Hyperthermophilic Archaeon Thermococcus kodakarensis"

    Article Title: Markerless Gene Editing in the Hyperthermophilic Archaeon Thermococcus kodakarensis

    Journal: Bio-protocol

    doi: 10.21769/BioProtoc.2604

    Diagnostic PCR confirms the deletion of TK0566 from the T. kodakarensis genome The same primer pairs are used on the TS559 genome as the deletion genome. The size shift using primers A/B corresponds to the deletion of TK0566 while the absence of products for the reactions using C/D and E/F confirms the gene has been deleted from its native locus. The absence of products using primers C/D demonstrates that TK0566 is not present anywhere in the genome of the deletion strain.
    Figure Legend Snippet: Diagnostic PCR confirms the deletion of TK0566 from the T. kodakarensis genome The same primer pairs are used on the TS559 genome as the deletion genome. The size shift using primers A/B corresponds to the deletion of TK0566 while the absence of products for the reactions using C/D and E/F confirms the gene has been deleted from its native locus. The absence of products using primers C/D demonstrates that TK0566 is not present anywhere in the genome of the deletion strain.

    Techniques Used: Diagnostic Assay, Polymerase Chain Reaction

    Overview of the markerless deletion scheme used in T. kodakarensis At the top of the figure is the B-plasmid used to delete the target gene from the genome. The plasmid recombines into the genome providing agmatine prototrophy to recipient cells and yields an intermediate genome. Two intermediate genomes are possible; however only one is depicted here. A second spontaneous recombination event excises plasmid sequences and permits survival in the presence of cytotoxic 6-MP. This second recombination event will result in the desired deletion genome (left) or the restoration of the TS559 genome (right).
    Figure Legend Snippet: Overview of the markerless deletion scheme used in T. kodakarensis At the top of the figure is the B-plasmid used to delete the target gene from the genome. The plasmid recombines into the genome providing agmatine prototrophy to recipient cells and yields an intermediate genome. Two intermediate genomes are possible; however only one is depicted here. A second spontaneous recombination event excises plasmid sequences and permits survival in the presence of cytotoxic 6-MP. This second recombination event will result in the desired deletion genome (left) or the restoration of the TS559 genome (right).

    Techniques Used: Plasmid Preparation

    Related Articles

    Clone Assay:

    Article Title: The Type II Secreted Lipase/Esterase LesA is a Key Virulence Factor Required for Xylella fastidiosa Pathogenesis in Grapevines
    Article Snippet: .. The insertion of cloned genes was verified by PCR using primers for sites flanking LesA genes, followed by transformation of ElectroMAX DH5α competent cells (Life Technologies, USA). .. For heterologous expression, cells were grown in LB medium supplemented with kanamycin (50 μg/mL) at 37 °C and 120 rpm to an A600 of 0.8.

    Article Title: Human Serum Albumin and p53-Activating Peptide Fusion Protein Is Able to Promote Apoptosis and Deliver Fatty Acid-Modified Molecules
    Article Snippet: .. Pichia pastoris yeast cells (Invitrogen, 18258-012 ) were then transformed using linearized pMM1 (for HSA-P53i) or pMM2 (for rHSA-PMI) plasmid DNA and rHSA-P53i and rHSA-PMI clones were selected for Zeocin resistance after 72 hours. .. Recombinant proteins were then expressed in Pichia pastoris according to the manufacturer’s instructions (Invitrogen, K1740-01 ).

    Article Title: Feline immunodeficiency virus (FIV) env recombinants are common in natural infections
    Article Snippet: .. Strain-specific primers for second round PCR incorporated restriction sites for subsequent cloning into the eukaryotic expression vector VR1012 [ ] and transformation into E. coli MAX Efficiency® DH5α™ Competent Cells (Invitrogen). .. Each VR1012-FIV env construct was sequenced using Big Dye Terminator v1.1 kit (Applied Biosystems) on an Applied Biosystems 3130xl capillary sequencer.

    Transfection:

    Article Title: In vivo Targeting of Adoptively Transferred T-cells with Antibody- and Cytokine-Conjugated Liposomes
    Article Snippet: .. DiD, ACK lysis buffer, Calcium Phosphate Transfection Kit, HEK293 Free Style Cells, Max Efficiency ® DH5α™ Competent cells and Phoenix eco viral packaging cells were obtained from Invitrogen Life Technologies (Grand Island, NY). .. Anti-thy1.1 (clone 19E12) and mouse IgG2a isotype control antibodies were purchased from BioXCell (West Lebanon, NH).

    other:

    Article Title: Cloning Should Be Simple: Escherichia coli DH5α-Mediated Assembly of Multiple DNA Fragments with Short End Homologies
    Article Snippet: Bacterial strains The following commercial products were used: Max Efficiency DH5α (Life Technologies, Carlsbad, CA; chemically competent, ~109 colony-forming unit or CFU / μg pUC19), High Efficiency NEB 5-alpha (New England Biolabs, Ipswich, MA; chemically competent, CFU ~109 /μg pUC19), and NEB 5-alpha Electrocompetent E . coli (New England Biolabs, CFU ~1010 /μg pUC19).

    Expressing:

    Article Title: Feline immunodeficiency virus (FIV) env recombinants are common in natural infections
    Article Snippet: .. Strain-specific primers for second round PCR incorporated restriction sites for subsequent cloning into the eukaryotic expression vector VR1012 [ ] and transformation into E. coli MAX Efficiency® DH5α™ Competent Cells (Invitrogen). .. Each VR1012-FIV env construct was sequenced using Big Dye Terminator v1.1 kit (Applied Biosystems) on an Applied Biosystems 3130xl capillary sequencer.

    Polymerase Chain Reaction:

    Article Title: The Type II Secreted Lipase/Esterase LesA is a Key Virulence Factor Required for Xylella fastidiosa Pathogenesis in Grapevines
    Article Snippet: .. The insertion of cloned genes was verified by PCR using primers for sites flanking LesA genes, followed by transformation of ElectroMAX DH5α competent cells (Life Technologies, USA). .. For heterologous expression, cells were grown in LB medium supplemented with kanamycin (50 μg/mL) at 37 °C and 120 rpm to an A600 of 0.8.

    Article Title: Feline immunodeficiency virus (FIV) env recombinants are common in natural infections
    Article Snippet: .. Strain-specific primers for second round PCR incorporated restriction sites for subsequent cloning into the eukaryotic expression vector VR1012 [ ] and transformation into E. coli MAX Efficiency® DH5α™ Competent Cells (Invitrogen). .. Each VR1012-FIV env construct was sequenced using Big Dye Terminator v1.1 kit (Applied Biosystems) on an Applied Biosystems 3130xl capillary sequencer.

    Article Title: Markerless Gene Editing in the Hyperthermophilic Archaeon Thermococcus kodakarensis
    Article Snippet: .. 1 ml TB syringe (BD, catalog number: 309624) 1.7 ml microcentrifuge tubes (VWR, catalog number: 490004-444) 0.2 ml PCR tubes (VWR, catalog number: 20170-012) Polystyrene Petri plates (Fisher Scientific, catalog number: S33580A) Split rubber stopper (DWK Life Sciences, Wheaton, catalog number: W224100-282) 20 mm aluminum seals (DWK Life Sciences, Wheaton, catalog number: 224178-01) 20 mm E-Z Crimper, Standard Seal (DWK Life Sciences, Wheaton, catalog number: W225303) 20 mm E-Z Decapper (DWK Life Sciences, Wheaton, catalog number: W225353) Polycarbonate centrifuge tubes (Beckman Coulter, catalog number: 361690) Cell spreader (Fisher Scientific, catalog number: 08-100-10) 10 ml serum bottles (DWK Life Sciences, Wheaton, catalog number: 223739) Face shields, lab coats, and autoclave gloves Paper towels T. kodakarensis strain TS559 ( ) DH5α E. coli competent cells (Thermo Fisher Scientific, Invitrogen™, catalog number: 18258012) XL1-Blue E. coli competent cells (Agilent Technologies, catalog number: 200228) pTS700 ( ) Note: Please contact corresponding author to obtain plasmid. .. Agmatine (Sigma-Aldrich, catalog number: A7127) Elemental sulfur (Aqua Solutions, catalog number: S8800-12KG) 10 mM Tris-HCl pH 8.0 (VWR, catalog number: 97061-258) Isopropanol (Sigma-Aldrich, catalog number: 190764) LE Quick dissolve agarose (VWR, catalog number: 490000-004) Ethidium bromide (Sigma-Aldrich, catalog number: E1510) AMPure XP (Fisher Scientific, catalog number: NC9959336) Manufacturer: Beckman Coulter, catalog number: .

    Lysis:

    Article Title: In vivo Targeting of Adoptively Transferred T-cells with Antibody- and Cytokine-Conjugated Liposomes
    Article Snippet: .. DiD, ACK lysis buffer, Calcium Phosphate Transfection Kit, HEK293 Free Style Cells, Max Efficiency ® DH5α™ Competent cells and Phoenix eco viral packaging cells were obtained from Invitrogen Life Technologies (Grand Island, NY). .. Anti-thy1.1 (clone 19E12) and mouse IgG2a isotype control antibodies were purchased from BioXCell (West Lebanon, NH).

    Transformation Assay:

    Article Title: The Type II Secreted Lipase/Esterase LesA is a Key Virulence Factor Required for Xylella fastidiosa Pathogenesis in Grapevines
    Article Snippet: .. The insertion of cloned genes was verified by PCR using primers for sites flanking LesA genes, followed by transformation of ElectroMAX DH5α competent cells (Life Technologies, USA). .. For heterologous expression, cells were grown in LB medium supplemented with kanamycin (50 μg/mL) at 37 °C and 120 rpm to an A600 of 0.8.

    Article Title: Human Serum Albumin and p53-Activating Peptide Fusion Protein Is Able to Promote Apoptosis and Deliver Fatty Acid-Modified Molecules
    Article Snippet: .. Pichia pastoris yeast cells (Invitrogen, 18258-012 ) were then transformed using linearized pMM1 (for HSA-P53i) or pMM2 (for rHSA-PMI) plasmid DNA and rHSA-P53i and rHSA-PMI clones were selected for Zeocin resistance after 72 hours. .. Recombinant proteins were then expressed in Pichia pastoris according to the manufacturer’s instructions (Invitrogen, K1740-01 ).

    Article Title: Feline immunodeficiency virus (FIV) env recombinants are common in natural infections
    Article Snippet: .. Strain-specific primers for second round PCR incorporated restriction sites for subsequent cloning into the eukaryotic expression vector VR1012 [ ] and transformation into E. coli MAX Efficiency® DH5α™ Competent Cells (Invitrogen). .. Each VR1012-FIV env construct was sequenced using Big Dye Terminator v1.1 kit (Applied Biosystems) on an Applied Biosystems 3130xl capillary sequencer.

    Article Title: The ATRX cDNA is prone to bacterial IS10 element insertions that alter its structure
    Article Snippet: .. ATRX transformation and growth in different bacterial strains ElectroMAX™ Stbl4™ competent cells (Life Technologies, 11635-018) and DH5α competent cells (Life Technologies 18258-012) were transformed according to manufacturer’s indications with 50 ng of IF-GFP-ATRX. .. Colonies from three independent transfections were picked and diluted in 100μl of PBS.

    Plasmid Preparation:

    Article Title: Human Serum Albumin and p53-Activating Peptide Fusion Protein Is Able to Promote Apoptosis and Deliver Fatty Acid-Modified Molecules
    Article Snippet: .. Pichia pastoris yeast cells (Invitrogen, 18258-012 ) were then transformed using linearized pMM1 (for HSA-P53i) or pMM2 (for rHSA-PMI) plasmid DNA and rHSA-P53i and rHSA-PMI clones were selected for Zeocin resistance after 72 hours. .. Recombinant proteins were then expressed in Pichia pastoris according to the manufacturer’s instructions (Invitrogen, K1740-01 ).

    Article Title: Feline immunodeficiency virus (FIV) env recombinants are common in natural infections
    Article Snippet: .. Strain-specific primers for second round PCR incorporated restriction sites for subsequent cloning into the eukaryotic expression vector VR1012 [ ] and transformation into E. coli MAX Efficiency® DH5α™ Competent Cells (Invitrogen). .. Each VR1012-FIV env construct was sequenced using Big Dye Terminator v1.1 kit (Applied Biosystems) on an Applied Biosystems 3130xl capillary sequencer.

    Article Title: Markerless Gene Editing in the Hyperthermophilic Archaeon Thermococcus kodakarensis
    Article Snippet: .. 1 ml TB syringe (BD, catalog number: 309624) 1.7 ml microcentrifuge tubes (VWR, catalog number: 490004-444) 0.2 ml PCR tubes (VWR, catalog number: 20170-012) Polystyrene Petri plates (Fisher Scientific, catalog number: S33580A) Split rubber stopper (DWK Life Sciences, Wheaton, catalog number: W224100-282) 20 mm aluminum seals (DWK Life Sciences, Wheaton, catalog number: 224178-01) 20 mm E-Z Crimper, Standard Seal (DWK Life Sciences, Wheaton, catalog number: W225303) 20 mm E-Z Decapper (DWK Life Sciences, Wheaton, catalog number: W225353) Polycarbonate centrifuge tubes (Beckman Coulter, catalog number: 361690) Cell spreader (Fisher Scientific, catalog number: 08-100-10) 10 ml serum bottles (DWK Life Sciences, Wheaton, catalog number: 223739) Face shields, lab coats, and autoclave gloves Paper towels T. kodakarensis strain TS559 ( ) DH5α E. coli competent cells (Thermo Fisher Scientific, Invitrogen™, catalog number: 18258012) XL1-Blue E. coli competent cells (Agilent Technologies, catalog number: 200228) pTS700 ( ) Note: Please contact corresponding author to obtain plasmid. .. Agmatine (Sigma-Aldrich, catalog number: A7127) Elemental sulfur (Aqua Solutions, catalog number: S8800-12KG) 10 mM Tris-HCl pH 8.0 (VWR, catalog number: 97061-258) Isopropanol (Sigma-Aldrich, catalog number: 190764) LE Quick dissolve agarose (VWR, catalog number: 490000-004) Ethidium bromide (Sigma-Aldrich, catalog number: E1510) AMPure XP (Fisher Scientific, catalog number: NC9959336) Manufacturer: Beckman Coulter, catalog number: .

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  • 96
    Thermo Fisher max efficiency dh5α competent cells
    Diagnostic PCR confirms the deletion of TK0566 from the T. <t>kodakarensis</t> genome The same primer pairs are used on the <t>TS559</t> genome as the deletion genome. The size shift using primers A/B corresponds to the deletion of TK0566 while the absence of products for the reactions using C/D and E/F confirms the gene has been deleted from its native locus. The absence of products using primers C/D demonstrates that TK0566 is not present anywhere in the genome of the deletion strain.
    Max Efficiency Dh5α Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/max efficiency dh5α competent cells/product/Thermo Fisher
    Average 96 stars, based on 65 article reviews
    Price from $9.99 to $1999.99
    max efficiency dh5α competent cells - by Bioz Stars, 2020-08
    96/100 stars
      Buy from Supplier

    85
    Thermo Fisher max efficiency dh5α t1r competent cells
    Diagnostic PCR confirms the deletion of TK0566 from the T. <t>kodakarensis</t> genome The same primer pairs are used on the <t>TS559</t> genome as the deletion genome. The size shift using primers A/B corresponds to the deletion of TK0566 while the absence of products for the reactions using C/D and E/F confirms the gene has been deleted from its native locus. The absence of products using primers C/D demonstrates that TK0566 is not present anywhere in the genome of the deletion strain.
    Max Efficiency Dh5α T1r Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/max efficiency dh5α t1r competent cells/product/Thermo Fisher
    Average 85 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    max efficiency dh5α t1r competent cells - by Bioz Stars, 2020-08
    85/100 stars
      Buy from Supplier

    Image Search Results


    Diagnostic PCR confirms the deletion of TK0566 from the T. kodakarensis genome The same primer pairs are used on the TS559 genome as the deletion genome. The size shift using primers A/B corresponds to the deletion of TK0566 while the absence of products for the reactions using C/D and E/F confirms the gene has been deleted from its native locus. The absence of products using primers C/D demonstrates that TK0566 is not present anywhere in the genome of the deletion strain.

    Journal: Bio-protocol

    Article Title: Markerless Gene Editing in the Hyperthermophilic Archaeon Thermococcus kodakarensis

    doi: 10.21769/BioProtoc.2604

    Figure Lengend Snippet: Diagnostic PCR confirms the deletion of TK0566 from the T. kodakarensis genome The same primer pairs are used on the TS559 genome as the deletion genome. The size shift using primers A/B corresponds to the deletion of TK0566 while the absence of products for the reactions using C/D and E/F confirms the gene has been deleted from its native locus. The absence of products using primers C/D demonstrates that TK0566 is not present anywhere in the genome of the deletion strain.

    Article Snippet: 1 ml TB syringe (BD, catalog number: 309624) 1.7 ml microcentrifuge tubes (VWR, catalog number: 490004-444) 0.2 ml PCR tubes (VWR, catalog number: 20170-012) Polystyrene Petri plates (Fisher Scientific, catalog number: S33580A) Split rubber stopper (DWK Life Sciences, Wheaton, catalog number: W224100-282) 20 mm aluminum seals (DWK Life Sciences, Wheaton, catalog number: 224178-01) 20 mm E-Z Crimper, Standard Seal (DWK Life Sciences, Wheaton, catalog number: W225303) 20 mm E-Z Decapper (DWK Life Sciences, Wheaton, catalog number: W225353) Polycarbonate centrifuge tubes (Beckman Coulter, catalog number: 361690) Cell spreader (Fisher Scientific, catalog number: 08-100-10) 10 ml serum bottles (DWK Life Sciences, Wheaton, catalog number: 223739) Face shields, lab coats, and autoclave gloves Paper towels T. kodakarensis strain TS559 ( ) DH5α E. coli competent cells (Thermo Fisher Scientific, Invitrogen™, catalog number: 18258012) XL1-Blue E. coli competent cells (Agilent Technologies, catalog number: 200228) pTS700 ( ) Note: Please contact corresponding author to obtain plasmid.

    Techniques: Diagnostic Assay, Polymerase Chain Reaction

    Overview of the markerless deletion scheme used in T. kodakarensis At the top of the figure is the B-plasmid used to delete the target gene from the genome. The plasmid recombines into the genome providing agmatine prototrophy to recipient cells and yields an intermediate genome. Two intermediate genomes are possible; however only one is depicted here. A second spontaneous recombination event excises plasmid sequences and permits survival in the presence of cytotoxic 6-MP. This second recombination event will result in the desired deletion genome (left) or the restoration of the TS559 genome (right).

    Journal: Bio-protocol

    Article Title: Markerless Gene Editing in the Hyperthermophilic Archaeon Thermococcus kodakarensis

    doi: 10.21769/BioProtoc.2604

    Figure Lengend Snippet: Overview of the markerless deletion scheme used in T. kodakarensis At the top of the figure is the B-plasmid used to delete the target gene from the genome. The plasmid recombines into the genome providing agmatine prototrophy to recipient cells and yields an intermediate genome. Two intermediate genomes are possible; however only one is depicted here. A second spontaneous recombination event excises plasmid sequences and permits survival in the presence of cytotoxic 6-MP. This second recombination event will result in the desired deletion genome (left) or the restoration of the TS559 genome (right).

    Article Snippet: 1 ml TB syringe (BD, catalog number: 309624) 1.7 ml microcentrifuge tubes (VWR, catalog number: 490004-444) 0.2 ml PCR tubes (VWR, catalog number: 20170-012) Polystyrene Petri plates (Fisher Scientific, catalog number: S33580A) Split rubber stopper (DWK Life Sciences, Wheaton, catalog number: W224100-282) 20 mm aluminum seals (DWK Life Sciences, Wheaton, catalog number: 224178-01) 20 mm E-Z Crimper, Standard Seal (DWK Life Sciences, Wheaton, catalog number: W225303) 20 mm E-Z Decapper (DWK Life Sciences, Wheaton, catalog number: W225353) Polycarbonate centrifuge tubes (Beckman Coulter, catalog number: 361690) Cell spreader (Fisher Scientific, catalog number: 08-100-10) 10 ml serum bottles (DWK Life Sciences, Wheaton, catalog number: 223739) Face shields, lab coats, and autoclave gloves Paper towels T. kodakarensis strain TS559 ( ) DH5α E. coli competent cells (Thermo Fisher Scientific, Invitrogen™, catalog number: 18258012) XL1-Blue E. coli competent cells (Agilent Technologies, catalog number: 200228) pTS700 ( ) Note: Please contact corresponding author to obtain plasmid.

    Techniques: Plasmid Preparation