t7 dna ligase  (New England Biolabs)


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  • 99
    Name:
    T7 DNA Ligase
    Description:
    T7 DNA Ligase 750 000 units
    Catalog Number:
    M0318L
    Price:
    276
    Category:
    DNA Ligases
    Size:
    750 000 units
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    Structured Review

    New England Biolabs t7 dna ligase
    T7 DNA Ligase
    T7 DNA Ligase 750 000 units
    https://www.bioz.com/result/t7 dna ligase/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t7 dna ligase - by Bioz Stars, 2021-05
    99/100 stars

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    Related Articles

    Ligation:

    Article Title: Improved transposon-based library preparation for the Ion Torrent platform
    Article Snippet: This pentaplet is complementary to the 5′-end of the exposed overhang and ensures ligation of the fragment to the oligonucleotide. .. The ligation reaction is carried out by T7 DNA ligase (New England BioLabs) to avoid blunt end ligation. .. The resulting library lacks complementary regions capable of forming hairpins; moreover, whereas only homodupexes have DNA strands equipped with both adaptors and are suitable for sequencing, heteroduplexes lack the second adaptor and do not participate in clonal amplifcation ( ).

    Article Title: Identification of focally amplified lineage-specific super-enhancers in human epithelial cancers
    Article Snippet: Cells stably expressing KRAB-dCas9 were then subsequently infected with sgRNAs and selected with 2μg/ml puromycin. .. For deletion of the e3 enhancer, tandem U6-promoter-sgRNA and H1-promoter-sgRNA cassettes were cloned into lentiCRISPR_v2 (Addgene #60954) for single vector expression of two sgRNAs as follows: 1) U6-sgRNA and H1-sgRNA products were generated by PCR amplification using the primers listed in , 2) PCR products were then digested with BsmBI to generate compatible sticky ends, 3) finally, three-way ligation of the two PCR products and BsmBI-digested lentiCRISPR_v2 was performed using T7 DNA ligase (NEB #M0318). .. Control ‘empty’ lentiCRISPR_v2 lacking expression of any sgRNAs was generated by BsmBI digestion, followed by blunting of ends (NEB #E1201) and ligation with T4 DNA ligase (NEB #M0202).

    CRISPR:

    Article Title: Hsp70 and NF-kB Mediated Control of Innate Inflammatory Responses in a Canine Macrophage Cell Line
    Article Snippet: Briefly, 5′ ends of gRNA oligos (10 μM) were phosphorylated and annealed with T4 PNK (New England Biolabs, Ipswich, UK) using the following cycle parameters: 30 min at 37 °C, 5 min at 95 °C, then ramp down to 25 °C at 0.3 °C/min and 4 °C. .. Subsequently, the annealed gRNAs were ligated into the CRISPR-concatemer vector using 100 ng CRISPR-concatemer vector, 10.0 μL oligo mixture, 1.0 μL BSA-containing restriction enzyme buffer (10×), 1.0 μL DTT (10 mM), 1.0 μL ATP (10 mM), 1.0 μL Bbs I, 1.0 μL T7 ligase, 5.0 μL H2 O, and the following cycle parameters: 25 cycles of 5 min at 37 °C and 5 min at 21 °C, hold for 15 min at 37 °C and then 4 °C forever. .. Ligated CRISPR-concatemer vectors were transformed into DH5α.

    Plasmid Preparation:

    Article Title: Hsp70 and NF-kB Mediated Control of Innate Inflammatory Responses in a Canine Macrophage Cell Line
    Article Snippet: Briefly, 5′ ends of gRNA oligos (10 μM) were phosphorylated and annealed with T4 PNK (New England Biolabs, Ipswich, UK) using the following cycle parameters: 30 min at 37 °C, 5 min at 95 °C, then ramp down to 25 °C at 0.3 °C/min and 4 °C. .. Subsequently, the annealed gRNAs were ligated into the CRISPR-concatemer vector using 100 ng CRISPR-concatemer vector, 10.0 μL oligo mixture, 1.0 μL BSA-containing restriction enzyme buffer (10×), 1.0 μL DTT (10 mM), 1.0 μL ATP (10 mM), 1.0 μL Bbs I, 1.0 μL T7 ligase, 5.0 μL H2 O, and the following cycle parameters: 25 cycles of 5 min at 37 °C and 5 min at 21 °C, hold for 15 min at 37 °C and then 4 °C forever. .. Ligated CRISPR-concatemer vectors were transformed into DH5α.

    Article Title: Biological Parts for Kluyveromyces marxianus Synthetic Biology
    Article Snippet: .. In a typical reaction, 40 fmol of each insert (either as an existing level I plasmid or PCR product) were combined with 20 fmol of the plasmid containing the backbone for the final product to be assembled along with 1μL T4 DNA ligase 0.5 μL each of T7 DNA ligase (3,000U μL−1 ) and BsmBI or BsaI, (10U μL−1 , NEB) and water to a total volume of 10 μL. .. For Golden Gate assembly the protocol was as follows: 25 cycles of digestion and ligation (2 min at 42°C followed by 5 min at 16°C), followed by 10 min digestion at 60°C and 10 min inactivation at 80°C.

    Article Title: Identification of focally amplified lineage-specific super-enhancers in human epithelial cancers
    Article Snippet: Cells stably expressing KRAB-dCas9 were then subsequently infected with sgRNAs and selected with 2μg/ml puromycin. .. For deletion of the e3 enhancer, tandem U6-promoter-sgRNA and H1-promoter-sgRNA cassettes were cloned into lentiCRISPR_v2 (Addgene #60954) for single vector expression of two sgRNAs as follows: 1) U6-sgRNA and H1-sgRNA products were generated by PCR amplification using the primers listed in , 2) PCR products were then digested with BsmBI to generate compatible sticky ends, 3) finally, three-way ligation of the two PCR products and BsmBI-digested lentiCRISPR_v2 was performed using T7 DNA ligase (NEB #M0318). .. Control ‘empty’ lentiCRISPR_v2 lacking expression of any sgRNAs was generated by BsmBI digestion, followed by blunting of ends (NEB #E1201) and ligation with T4 DNA ligase (NEB #M0202).

    Article Title: Rapid characterization of spike variants via mammalian cell surface display
    Article Snippet: Golden Gate compatible parts were arranged in a 384-well Echo Source Plate (PP-0200) and transferred to 96-well PCR destination plates using the Echo 525. .. Each well of the 96-well destination plate received the following Golden Gate reaction mixture: 0.5 μL of T7 DNA Ligase (NEB; M0318S), 0.5 μL of AarI (Thermo Fisher; ER1582), 0.2 μL AarI Oligo (Thermo Fisher), 1 μL T4 DNA Ligase Buffer (NEB; B0202A), 1 μL of the insert and plasmid DNA, and nuclease-free water to bring the final volume to 10 μL per reaction. .. Reaction mixtures were incubated on a thermocycler according to the following conditions: 25 cycles of digestion and ligation (37 °C for 1 min, and 16 °C for 2 mins), followed by a final digestion (37 °C for 30 mins), and a heat inactivation step (80 °C for 20 mins).

    Article Title: Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
    Article Snippet: The PCR products were purified and then digested for 2 hrs at 37°C with Aar I (Thermo Fisher Scientific). .. A Level 1 Acceptor Vector A was also digested with AarI and ligated to the Auxiliary Plasmid cassettes with 1 μl T7 DNA ligase (NEB) for 20min at RT. .. The constructs were verified by Sanger sequencing (GATC-Biotech or Edinburgh Genomics).

    Polymerase Chain Reaction:

    Article Title: Biological Parts for Kluyveromyces marxianus Synthetic Biology
    Article Snippet: .. In a typical reaction, 40 fmol of each insert (either as an existing level I plasmid or PCR product) were combined with 20 fmol of the plasmid containing the backbone for the final product to be assembled along with 1μL T4 DNA ligase 0.5 μL each of T7 DNA ligase (3,000U μL−1 ) and BsmBI or BsaI, (10U μL−1 , NEB) and water to a total volume of 10 μL. .. For Golden Gate assembly the protocol was as follows: 25 cycles of digestion and ligation (2 min at 42°C followed by 5 min at 16°C), followed by 10 min digestion at 60°C and 10 min inactivation at 80°C.

    Article Title: Identification of focally amplified lineage-specific super-enhancers in human epithelial cancers
    Article Snippet: Cells stably expressing KRAB-dCas9 were then subsequently infected with sgRNAs and selected with 2μg/ml puromycin. .. For deletion of the e3 enhancer, tandem U6-promoter-sgRNA and H1-promoter-sgRNA cassettes were cloned into lentiCRISPR_v2 (Addgene #60954) for single vector expression of two sgRNAs as follows: 1) U6-sgRNA and H1-sgRNA products were generated by PCR amplification using the primers listed in , 2) PCR products were then digested with BsmBI to generate compatible sticky ends, 3) finally, three-way ligation of the two PCR products and BsmBI-digested lentiCRISPR_v2 was performed using T7 DNA ligase (NEB #M0318). .. Control ‘empty’ lentiCRISPR_v2 lacking expression of any sgRNAs was generated by BsmBI digestion, followed by blunting of ends (NEB #E1201) and ligation with T4 DNA ligase (NEB #M0202).

    Clone Assay:

    Article Title: Identification of focally amplified lineage-specific super-enhancers in human epithelial cancers
    Article Snippet: Cells stably expressing KRAB-dCas9 were then subsequently infected with sgRNAs and selected with 2μg/ml puromycin. .. For deletion of the e3 enhancer, tandem U6-promoter-sgRNA and H1-promoter-sgRNA cassettes were cloned into lentiCRISPR_v2 (Addgene #60954) for single vector expression of two sgRNAs as follows: 1) U6-sgRNA and H1-sgRNA products were generated by PCR amplification using the primers listed in , 2) PCR products were then digested with BsmBI to generate compatible sticky ends, 3) finally, three-way ligation of the two PCR products and BsmBI-digested lentiCRISPR_v2 was performed using T7 DNA ligase (NEB #M0318). .. Control ‘empty’ lentiCRISPR_v2 lacking expression of any sgRNAs was generated by BsmBI digestion, followed by blunting of ends (NEB #E1201) and ligation with T4 DNA ligase (NEB #M0202).

    Expressing:

    Article Title: Identification of focally amplified lineage-specific super-enhancers in human epithelial cancers
    Article Snippet: Cells stably expressing KRAB-dCas9 were then subsequently infected with sgRNAs and selected with 2μg/ml puromycin. .. For deletion of the e3 enhancer, tandem U6-promoter-sgRNA and H1-promoter-sgRNA cassettes were cloned into lentiCRISPR_v2 (Addgene #60954) for single vector expression of two sgRNAs as follows: 1) U6-sgRNA and H1-sgRNA products were generated by PCR amplification using the primers listed in , 2) PCR products were then digested with BsmBI to generate compatible sticky ends, 3) finally, three-way ligation of the two PCR products and BsmBI-digested lentiCRISPR_v2 was performed using T7 DNA ligase (NEB #M0318). .. Control ‘empty’ lentiCRISPR_v2 lacking expression of any sgRNAs was generated by BsmBI digestion, followed by blunting of ends (NEB #E1201) and ligation with T4 DNA ligase (NEB #M0202).

    Generated:

    Article Title: Identification of focally amplified lineage-specific super-enhancers in human epithelial cancers
    Article Snippet: Cells stably expressing KRAB-dCas9 were then subsequently infected with sgRNAs and selected with 2μg/ml puromycin. .. For deletion of the e3 enhancer, tandem U6-promoter-sgRNA and H1-promoter-sgRNA cassettes were cloned into lentiCRISPR_v2 (Addgene #60954) for single vector expression of two sgRNAs as follows: 1) U6-sgRNA and H1-sgRNA products were generated by PCR amplification using the primers listed in , 2) PCR products were then digested with BsmBI to generate compatible sticky ends, 3) finally, three-way ligation of the two PCR products and BsmBI-digested lentiCRISPR_v2 was performed using T7 DNA ligase (NEB #M0318). .. Control ‘empty’ lentiCRISPR_v2 lacking expression of any sgRNAs was generated by BsmBI digestion, followed by blunting of ends (NEB #E1201) and ligation with T4 DNA ligase (NEB #M0202).

    Amplification:

    Article Title: Identification of focally amplified lineage-specific super-enhancers in human epithelial cancers
    Article Snippet: Cells stably expressing KRAB-dCas9 were then subsequently infected with sgRNAs and selected with 2μg/ml puromycin. .. For deletion of the e3 enhancer, tandem U6-promoter-sgRNA and H1-promoter-sgRNA cassettes were cloned into lentiCRISPR_v2 (Addgene #60954) for single vector expression of two sgRNAs as follows: 1) U6-sgRNA and H1-sgRNA products were generated by PCR amplification using the primers listed in , 2) PCR products were then digested with BsmBI to generate compatible sticky ends, 3) finally, three-way ligation of the two PCR products and BsmBI-digested lentiCRISPR_v2 was performed using T7 DNA ligase (NEB #M0318). .. Control ‘empty’ lentiCRISPR_v2 lacking expression of any sgRNAs was generated by BsmBI digestion, followed by blunting of ends (NEB #E1201) and ligation with T4 DNA ligase (NEB #M0202).

    Construct:

    Article Title: Fronto-temporal dementia risk gene TMEM106B has opposing effects in different lysosomal storage disorders
    Article Snippet: .. To generate sgRNA-expressing constructs, pX552 was digested using SapI Fast Digest (ThermoFisher Scientific, D1934) and annealed oligos were ligated using T7 DNA Ligase (NEB, M0318). .. Transformation was performed using One-Shot Stbl3 Chemically Competent Escherichia coli (Thermo, 737303).

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  • 99
    New England Biolabs t7 dna ligase
    T7 Dna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 dna ligase/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t7 dna ligase - by Bioz Stars, 2021-05
    99/100 stars
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