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Lucigen Corp t4 ligase
T4 Ligase, supplied by Lucigen Corp, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/t4 ligase/product/Lucigen Corp
Average 92 stars, based on 1 article reviews
Price from $9.99 to $1999.99
t4 ligase - by Bioz Stars, 2020-05
92/100 stars

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Article Title: K+ Channel Facilitation of Exocytosis by Dynamic Interaction with Syntaxin
Article Snippet: Restriction enzymes were purchased from New England Biolabs (Ipswich, MA), and T4 ligase was purchased from Lucigen Corporation (Middleton, WI).

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    Lucigen Corp t4 dna ligase
    Strategy for PCR-free cloning of large bacterial genetic regions. The major steps of cloning large genetic inserts are indicated. The catR-oriT-oriR (pMJH97) cassette was PCR amplified using primer pairs with 50–60 bp homologous sequence at their 5′-ends specific to the targetred site. Depending on the choice of restriction enzymes, the resulting dsDNA substrate can be integrated upstream or downstream of the targeted site of the genome using the recombineering system. Once the catR-oriT-oriR (pMJH97) cassette integration into the genome was confirmed by PCR and sequencing using primers P1 and P2, the genomic DNA of integrants was restriction digested with an appropriate restriction enzyme to clone into E. coli after self-ligation using <t>T4</t> DNA ligase. The cloning of the correct insert into the plasmid pMJH97 was verified by PCR and sequencing using vector and insert specific primers P3 and P4, respectively. The plasmids with cloned inserts were then readily transfered to other Gram-negative bacterial strain by oriT sequence-mediated conjugal transfer using an appropriate donor strain.
    T4 Dna Ligase, supplied by Lucigen Corp, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 dna ligase/product/Lucigen Corp
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t4 dna ligase - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    92
    Lucigen Corp t4 ligase
    Strategy for PCR-free cloning of large bacterial genetic regions. The major steps of cloning large genetic inserts are indicated. The catR-oriT-oriR (pMJH97) cassette was PCR amplified using primer pairs with 50–60 bp homologous sequence at their 5′-ends specific to the targetred site. Depending on the choice of restriction enzymes, the resulting dsDNA substrate can be integrated upstream or downstream of the targeted site of the genome using the recombineering system. Once the catR-oriT-oriR (pMJH97) cassette integration into the genome was confirmed by PCR and sequencing using primers P1 and P2, the genomic DNA of integrants was restriction digested with an appropriate restriction enzyme to clone into E. coli after self-ligation using <t>T4</t> DNA ligase. The cloning of the correct insert into the plasmid pMJH97 was verified by PCR and sequencing using vector and insert specific primers P3 and P4, respectively. The plasmids with cloned inserts were then readily transfered to other Gram-negative bacterial strain by oriT sequence-mediated conjugal transfer using an appropriate donor strain.
    T4 Ligase, supplied by Lucigen Corp, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 ligase/product/Lucigen Corp
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t4 ligase - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

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    Strategy for PCR-free cloning of large bacterial genetic regions. The major steps of cloning large genetic inserts are indicated. The catR-oriT-oriR (pMJH97) cassette was PCR amplified using primer pairs with 50–60 bp homologous sequence at their 5′-ends specific to the targetred site. Depending on the choice of restriction enzymes, the resulting dsDNA substrate can be integrated upstream or downstream of the targeted site of the genome using the recombineering system. Once the catR-oriT-oriR (pMJH97) cassette integration into the genome was confirmed by PCR and sequencing using primers P1 and P2, the genomic DNA of integrants was restriction digested with an appropriate restriction enzyme to clone into E. coli after self-ligation using T4 DNA ligase. The cloning of the correct insert into the plasmid pMJH97 was verified by PCR and sequencing using vector and insert specific primers P3 and P4, respectively. The plasmids with cloned inserts were then readily transfered to other Gram-negative bacterial strain by oriT sequence-mediated conjugal transfer using an appropriate donor strain.

    Journal: Biotechnology Reports

    Article Title: Genome modifications and cloning using a conjugally transferable recombineering system

    doi: 10.1016/j.btre.2015.08.005

    Figure Lengend Snippet: Strategy for PCR-free cloning of large bacterial genetic regions. The major steps of cloning large genetic inserts are indicated. The catR-oriT-oriR (pMJH97) cassette was PCR amplified using primer pairs with 50–60 bp homologous sequence at their 5′-ends specific to the targetred site. Depending on the choice of restriction enzymes, the resulting dsDNA substrate can be integrated upstream or downstream of the targeted site of the genome using the recombineering system. Once the catR-oriT-oriR (pMJH97) cassette integration into the genome was confirmed by PCR and sequencing using primers P1 and P2, the genomic DNA of integrants was restriction digested with an appropriate restriction enzyme to clone into E. coli after self-ligation using T4 DNA ligase. The cloning of the correct insert into the plasmid pMJH97 was verified by PCR and sequencing using vector and insert specific primers P3 and P4, respectively. The plasmids with cloned inserts were then readily transfered to other Gram-negative bacterial strain by oriT sequence-mediated conjugal transfer using an appropriate donor strain.

    Article Snippet: Blunt-ended and purified genomic DNA fragments were self-ligated using T4 DNA ligase and electroporated into E. coli (E. cloni 10G, Lucigen Corp.) for cloning.

    Techniques: Polymerase Chain Reaction, Clone Assay, Amplification, Sequencing, Ligation, Plasmid Preparation