taq dna ligase  (New England Biolabs)


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    Name:
    Taq DNA Ligase
    Description:
    Taq DNA Ligase 10 000 units
    Catalog Number:
    m0208l
    Price:
    316
    Size:
    10 000 units
    Category:
    DNA Ligases
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    Structured Review

    New England Biolabs taq dna ligase
    Taq DNA Ligase
    Taq DNA Ligase 10 000 units
    https://www.bioz.com/result/taq dna ligase/product/New England Biolabs
    Average 90 stars, based on 631 article reviews
    Price from $9.99 to $1999.99
    taq dna ligase - by Bioz Stars, 2020-01
    90/100 stars

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    Related Articles

    Transduction:

    Article Title: RemA is a DNA-binding protein that activates biofilm matrix gene expression in Bacillus subtilis
    Article Snippet: The amplified product was transformed into competent cells of PY79 and then transferred to the 3610 background using SPP1-mediated generalized transduction. .. An assembly master mixture was made by combining prepared 5× isothermal assembly reaction buffer (131 mM Tris-HCl, 13.1 mM MgCl2 , 13.1 mM DTT, 8.21 mM PEG-8000, 1.32 mM NAD and 0.26 mM each dNTP) with Phusion DNA polymerase (New England BioLabs) (0.033 units µl−1 ), T5 exonuclease diluted 1:5 with 5× reaction buffer (New England BioLabs) (0.01 units µl−1 ), Taq DNA ligase (New England BioLabs) (5328 units µl−1 ) and additional dNTPs (267 µM).

    Clone Assay:

    Article Title: Identifying components required for OMP biogenesis as novel targets for antiinfective drugs
    Article Snippet: The vector pSB890Y and the flanking regions were then assembled using a selfmade Gibson Assembly Master Mix containing T5 Exonuclease (10U µl, Epicentre), Phusion Polymerase (2U µl, New England Biolabs) and Taq-DNA-Ligase (40U µl, New England Biolabs). .. TetR clones (indicative for the presence of pSB890Y) were selected and the presence of the inserts comprising the up- and downstream regions of the target genes was verified by PCR with primers pSB890_seq_f and pSB890_seq_r.

    Article Title: PARIS, an optogenetic method for functionally mapping gap junctions
    Article Snippet: The fragments were assembled using T5 exonuclease, Phusion DNA polymerase, and Taq DNA ligase (New England Biolabs). .. For cultured cell expression experiments, genes were cloned into the pcDNA3.1 vector unless otherwise noted.

    Article Title: Engineering Corynebacterium glutamicum to produce 5-aminolevulinic acid from glucose
    Article Snippet: The three amplicons were ligated to pUC19, which was digested with Sal I (Fermentas, China) according to the method of Gibson assembly that included T5 exonuclease (Epicentre, USA), Phusion DNA polymerase (New England Biolabs, USA), and Taq DNA ligase (New England Biolabs, USA) [ ]. .. Moreover, gltX , hemA , and hemL were cloned from the genome of C. glutamicum ATCC 13032 using the primers cggltx-F, cggltx-R and cghema-F, cgdhema-R, and cgdheml-F, cgheml-R. To enhance the stability of HemA, hemA was cloned from the S. arizona genome, which encodes the same predicted HemA amino acid sequence of S. typhimurium .

    Amplification:

    Article Title: Association of the MicroRNA-146a SNP rs2910164 with Ischemic Stroke Incidence and Prognosis in a Chinese Population
    Article Snippet: .. Further amplification was conducted in 10-μL multiplex LDR reaction mixtures containing 100 ng of original PCR product (1 µL), 0.05 μL of NEB Taq DNA ligase ,1 μL of probe mix, and 6.95 μL of ddH2 O, using the ABI PRISM 377 DNA Sequencer detected and GeneMapper software (ABI) analyzed. ..

    Article Title: Identifying components required for OMP biogenesis as novel targets for antiinfective drugs
    Article Snippet: Also the flanking regions ∼1000 bp up- and downstream of the target genes were amplified by PCR using the primer pairs listed in the category “Generation of deletion mutants” in Table S2. .. The vector pSB890Y and the flanking regions were then assembled using a selfmade Gibson Assembly Master Mix containing T5 Exonuclease (10U µl, Epicentre), Phusion Polymerase (2U µl, New England Biolabs) and Taq-DNA-Ligase (40U µl, New England Biolabs).

    Article Title: The Human Myotrophin Variant Attenuates MicroRNA-Let-7 Binding Ability but Not Risk of Left Ventricular Hypertrophy in Human Essential Hypertension
    Article Snippet: .. Further amplification was performed in a 10 μl volume of multiplex LDR reaction mixture, containing 1 μl (100 ng) of the resultant probe mix, 1 μl of probe mix, 0.05 μl of NEB Taq DNA ligase and 7.95 μl of ddH2 O. ..

    Article Title: RemA is a DNA-binding protein that activates biofilm matrix gene expression in Bacillus subtilis
    Article Snippet: Insertions were verified by PCR amplification using primers 3318/3321. .. An assembly master mixture was made by combining prepared 5× isothermal assembly reaction buffer (131 mM Tris-HCl, 13.1 mM MgCl2 , 13.1 mM DTT, 8.21 mM PEG-8000, 1.32 mM NAD and 0.26 mM each dNTP) with Phusion DNA polymerase (New England BioLabs) (0.033 units µl−1 ), T5 exonuclease diluted 1:5 with 5× reaction buffer (New England BioLabs) (0.01 units µl−1 ), Taq DNA ligase (New England BioLabs) (5328 units µl−1 ) and additional dNTPs (267 µM).

    Article Title: Association of RAGE gene four single nucleotide polymorphisms with the risk, invasion, metastasis and overall survival of gastric cancer in Chinese
    Article Snippet: The steps were as follows: initial denaturation step at 94℃ for 3 min, amplification was carried out by 35 cycles at 94℃ for 15 s, 54℃ for 15 s, and at 72℃ for 30 s, followed by a final elongation cycle at 72℃ for 3 min. .. The connection reaction was performed at adding the PCR product 3 μl, 1 U 10×Taq DNA ligase buffer (New England Biolabs, United States), 0.125 μl Taq DNA ligase (40 U/μl, New England Biolabs, United States ), probe (10p) 0.01 μl/strip and with H2 O added to 10 μl under the condition of by cycles at 94℃ for 30 s, 56℃ for 3 min. LDR fluorescent products were analyzed by ABI 377 sequence analyzer (Perkin-Elmer, Foster City, CA, USA), and the results were analyzed by the fragment analysis software GeneMapper 3.7 (Applied Biosystems, Foster city, CA, USA).

    Article Title: Copy number variations exploration of multiple genes in Graves’ disease
    Article Snippet: The target DNA sequences were amplified using a multiplex PCR method, and the PCR program (20 μL volume) was conducted as follows: 95°C 2 minutes, 94°C 30 s, 40 cycles (53°C 90 s, 65°C 30 s), 65°C 10 minutes. .. The ligation reaction for each sample was carried out in a final volume of 10 μL, containing 1 μL of 1× buffer, 4 μL of multi-PCR product, 1 μL of 2 pmol/μl probe mix, and 0.05 μL of 20U Taq DNA ligase (New England Biolabs).

    Article Title: Polymorphism in the Vesicular Monoamine Transporter 2 Gene Decreases the Risk of Parkinson's Disease in Han Chinese Men
    Article Snippet: After amplification, 1 μ L of proteinase K (20 mg/mL) was added and the reactions were heated at 70°C for 10 min and then quenched at 94°C for 15 min. .. The ligation reaction for each subject was carried out in a final volume of 20 μ L containing 20 mM Tris-HCl (pH 7.6), 25 mM potassium acetate, 10 mM magnesium acetate, 10 mM DTT, 1 mM NAD, 0.1% Triton X-100, 10 μ L of multiplex PCR product, 1 pmol of each discriminating oligo, 1 pmol of each common oligo, and 0.5 μ L of 40 U/μ L Taq DNA ligase (New England Biolabs, USA).

    Article Title: Engineering Corynebacterium glutamicum to produce 5-aminolevulinic acid from glucose
    Article Snippet: The gltX , hemA , and hemL genes were amplified from the wild-type C. glutamicum ATCC 13032 genome using primers gltx-F, gltx-R and hema-F, hema-R, and heml-F, and heml-R, respectively. .. The three amplicons were ligated to pUC19, which was digested with Sal I (Fermentas, China) according to the method of Gibson assembly that included T5 exonuclease (Epicentre, USA), Phusion DNA polymerase (New England Biolabs, USA), and Taq DNA ligase (New England Biolabs, USA) [ ].

    Synthesized:

    Article Title: PARIS, an optogenetic method for functionally mapping gap junctions
    Article Snippet: The fragments were assembled using T5 exonuclease, Phusion DNA polymerase, and Taq DNA ligase (New England Biolabs). .. ArchT was amplified from pAAV-CAG-ArchT-GFP , codon-optimized rhodopsin genes from different species were synthesized by Qinglan Biotec then fused at the C-terminus with BFP2, a trafficking sequence (TS), and an ER export sequence (ERex), producing Actuator-BFP2.

    Cytometry:

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Detection of doubly labeled ligation products occurs through dual fluorescence flow cytometry in the BioPlex array reader (Bio-Rad Laboratories, Hercules, CA) and leads to collection of "reporter" signal in unique allele-specific bins. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Construct:

    Article Title: PARIS, an optogenetic method for functionally mapping gap junctions
    Article Snippet: Molecular cloning All plasmids were constructed using the Gibson assembly ( ) method. .. The fragments were assembled using T5 exonuclease, Phusion DNA polymerase, and Taq DNA ligase (New England Biolabs).

    Real-time Polymerase Chain Reaction:

    Article Title: Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus
    Article Snippet: NC-DNA (108 copies) was incubated with 32 units (U) of Thermostable FEN1 in ThermoPol Buffer (New England Biolabs) at 65°C for 10 min, followed by incubation with 8 U of Bst DNA polymerase, 40 U of Taq DNA ligase, 100 μM dNTPs, and NAD+ (all from New England Biolabs). .. After further incubation at 37°C for 20 min, DNA was purified by phenol/chloroform extraction and ethanol precipitation, and subjected to cccDNA-selective qPCR or RCA, as described above.

    Incubation:

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Following this hybridization reaction, products are incubated (Step #3) in a solution containing streptavidin-R-phycoerythrin (SA-PE) to allow "reporter" labeling through binding to the 3'-biotin on the conserved sequence primers. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Article Title: Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus
    Article Snippet: .. NC-DNA (108 copies) was incubated with 32 units (U) of Thermostable FEN1 in ThermoPol Buffer (New England Biolabs) at 65°C for 10 min, followed by incubation with 8 U of Bst DNA polymerase, 40 U of Taq DNA ligase, 100 μM dNTPs, and NAD+ (all from New England Biolabs). .. After further incubation at 37°C for 20 min, DNA was purified by phenol/chloroform extraction and ethanol precipitation, and subjected to cccDNA-selective qPCR or RCA, as described above.

    Article Title: Extensive sequencing of seven human genomes to characterize benchmark reference materials
    Article Snippet: .. Nick-labeled DNA was then incubated for 40 min at 37 °C using BioNano Genomics IrysPrep Repair mix and New England BioLabs Taq DNA Ligase at a final concentration of 1 U/μl. .. Labeled-repaired DNA was then recovered from the thin layer agarose by digesting with GELaseand counterstained with BioNano Genomics IrysPrep DNA Stain prior to data collection on the Irys system.

    Expressing:

    Article Title: PARIS, an optogenetic method for functionally mapping gap junctions
    Article Snippet: The fragments were assembled using T5 exonuclease, Phusion DNA polymerase, and Taq DNA ligase (New England Biolabs). .. For cultured cell expression experiments, genes were cloned into the pcDNA3.1 vector unless otherwise noted.

    Transformation Assay:

    Article Title: Identifying components required for OMP biogenesis as novel targets for antiinfective drugs
    Article Snippet: The vector pSB890Y and the flanking regions were then assembled using a selfmade Gibson Assembly Master Mix containing T5 Exonuclease (10U µl, Epicentre), Phusion Polymerase (2U µl, New England Biolabs) and Taq-DNA-Ligase (40U µl, New England Biolabs). .. Next, the strain E. coli 118λpir was transformed with the resulting plasmid.

    Article Title: RemA is a DNA-binding protein that activates biofilm matrix gene expression in Bacillus subtilis
    Article Snippet: The amplified product was transformed into competent cells of PY79 and then transferred to the 3610 background using SPP1-mediated generalized transduction. .. An assembly master mixture was made by combining prepared 5× isothermal assembly reaction buffer (131 mM Tris-HCl, 13.1 mM MgCl2 , 13.1 mM DTT, 8.21 mM PEG-8000, 1.32 mM NAD and 0.26 mM each dNTP) with Phusion DNA polymerase (New England BioLabs) (0.033 units µl−1 ), T5 exonuclease diluted 1:5 with 5× reaction buffer (New England BioLabs) (0.01 units µl−1 ), Taq DNA ligase (New England BioLabs) (5328 units µl−1 ) and additional dNTPs (267 µM).

    Hybridization:

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Following this hybridization reaction, products are incubated (Step #3) in a solution containing streptavidin-R-phycoerythrin (SA-PE) to allow "reporter" labeling through binding to the 3'-biotin on the conserved sequence primers. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Flow Cytometry:

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Detection of doubly labeled ligation products occurs through dual fluorescence flow cytometry in the BioPlex array reader (Bio-Rad Laboratories, Hercules, CA) and leads to collection of "reporter" signal in unique allele-specific bins. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Article Title: Integrative Detection and Analysis of Structural Variation in Cancer Genomes
    Article Snippet: Nick-labeled DNA was repaired in 1X repair mix (BioNano Genomics), 1× Thermo polymerase buffer (NEB), 50uM NAD+ (New England Biolabs), and 3ul 120U Taq DNA ligase (New England Biolabs) at 37°C for 30 minutes. .. DNA staining was finally performed with the final solution containing 1× flow buffer, 1× DTT (BioNano Genomics), and 3ul DNA stain (BioNano Genomics), in room temperature overnight.

    Ligation:

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Detection of doubly labeled ligation products occurs through dual fluorescence flow cytometry in the BioPlex array reader (Bio-Rad Laboratories, Hercules, CA) and leads to collection of "reporter" signal in unique allele-specific bins. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Article Title: Copy number variations exploration of multiple genes in Graves’ disease
    Article Snippet: .. The ligation reaction for each sample was carried out in a final volume of 10 μL, containing 1 μL of 1× buffer, 4 μL of multi-PCR product, 1 μL of 2 pmol/μl probe mix, and 0.05 μL of 20U Taq DNA ligase (New England Biolabs). ..

    Article Title: Polymorphism in the Vesicular Monoamine Transporter 2 Gene Decreases the Risk of Parkinson's Disease in Han Chinese Men
    Article Snippet: .. The ligation reaction for each subject was carried out in a final volume of 20 μ L containing 20 mM Tris-HCl (pH 7.6), 25 mM potassium acetate, 10 mM magnesium acetate, 10 mM DTT, 1 mM NAD, 0.1% Triton X-100, 10 μ L of multiplex PCR product, 1 pmol of each discriminating oligo, 1 pmol of each common oligo, and 0.5 μ L of 40 U/μ L Taq DNA ligase (New England Biolabs, USA). ..

    Cell Culture:

    Article Title: PARIS, an optogenetic method for functionally mapping gap junctions
    Article Snippet: The fragments were assembled using T5 exonuclease, Phusion DNA polymerase, and Taq DNA ligase (New England Biolabs). .. For cultured cell expression experiments, genes were cloned into the pcDNA3.1 vector unless otherwise noted.

    Article Title: Extensive sequencing of seven human genomes to characterize benchmark reference materials
    Article Snippet: Nick-labeled DNA was then incubated for 40 min at 37 °C using BioNano Genomics IrysPrep Repair mix and New England BioLabs Taq DNA Ligase at a final concentration of 1 U/μl. .. DNA was isolated from a lymphoid-cell cell culture of the Chinese son (GM24631) using the Bio-Rad CHEF Mammalian Genomic DNA Plug Kit protocol and lysed using BioNano Genomics IrysPrep Lysis Buffer and digested with QIAGEN Puregene Proteinase K. DNA was solubilized using GELase Agarose Gel-Digesting Preparation and drop-dialyzed before labeling using standard IrysPrep Reagent Kit protocols.

    Generated:

    Article Title: Identifying components required for OMP biogenesis as novel targets for antiinfective drugs
    Article Snippet: Generation of knockout strains The knockout strains, that are listed in Table S1 were generated using the suicide plasmid pSB890Y (a derivative of pSB890 where internal PstI restriction sites were removed). .. The vector pSB890Y and the flanking regions were then assembled using a selfmade Gibson Assembly Master Mix containing T5 Exonuclease (10U µl, Epicentre), Phusion Polymerase (2U µl, New England Biolabs) and Taq-DNA-Ligase (40U µl, New England Biolabs).

    Article Title: Biosensor libraries harness large classes of binding domains for construction of allosteric transcriptional regulators
    Article Snippet: This mock library enabled the testing of three different thermostable ligases: Ampligase (Epicentre, Madison, WI), 9°N DNA ligase (NEB, Ipswich, MA) and Taq DNA ligase (NEB, Ipswich, MA). .. The only chimeras generated through a canonical LCR were Class I.

    Sequencing:

    Article Title: The Human Myotrophin Variant Attenuates MicroRNA-Let-7 Binding Ability but Not Risk of Left Ventricular Hypertrophy in Human Essential Hypertension
    Article Snippet: Further amplification was performed in a 10 μl volume of multiplex LDR reaction mixture, containing 1 μl (100 ng) of the resultant probe mix, 1 μl of probe mix, 0.05 μl of NEB Taq DNA ligase and 7.95 μl of ddH2 O. .. Reproducibility of genotyping was confirmed by sequencing in 400 randomly selected samples with 100% concordance.

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Following this hybridization reaction, products are incubated (Step #3) in a solution containing streptavidin-R-phycoerythrin (SA-PE) to allow "reporter" labeling through binding to the 3'-biotin on the conserved sequence primers. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Article Title: Association of RAGE gene four single nucleotide polymorphisms with the risk, invasion, metastasis and overall survival of gastric cancer in Chinese
    Article Snippet: .. The connection reaction was performed at adding the PCR product 3 μl, 1 U 10×Taq DNA ligase buffer (New England Biolabs, United States), 0.125 μl Taq DNA ligase (40 U/μl, New England Biolabs, United States ), probe (10p) 0.01 μl/strip and with H2 O added to 10 μl under the condition of by cycles at 94℃ for 30 s, 56℃ for 3 min. LDR fluorescent products were analyzed by ABI 377 sequence analyzer (Perkin-Elmer, Foster City, CA, USA), and the results were analyzed by the fragment analysis software GeneMapper 3.7 (Applied Biosystems, Foster city, CA, USA). .. To test the accuracy of PCR-LDR method, a total of 96 DNA samples were randomly selected from the total sample, and they were genotyped using the same method, with exactly the sample findings for duplicated samples.

    Article Title: Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus
    Article Snippet: NC-DNA (108 copies) was incubated with 32 units (U) of Thermostable FEN1 in ThermoPol Buffer (New England Biolabs) at 65°C for 10 min, followed by incubation with 8 U of Bst DNA polymerase, 40 U of Taq DNA ligase, 100 μM dNTPs, and NAD+ (all from New England Biolabs). .. The sequence corresponding to gap region of rcDNA was confirmed by direct sequencing (oligonucleotide is listed in ).

    Article Title: PARIS, an optogenetic method for functionally mapping gap junctions
    Article Snippet: The fragments were assembled using T5 exonuclease, Phusion DNA polymerase, and Taq DNA ligase (New England Biolabs). .. All sequences were verified using Sanger sequencing in our in-house facility (sequencing platform in the School of Life Sciences of the Peking University).

    Binding Assay:

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Following this hybridization reaction, products are incubated (Step #3) in a solution containing streptavidin-R-phycoerythrin (SA-PE) to allow "reporter" labeling through binding to the 3'-biotin on the conserved sequence primers. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Molecular Cloning:

    Article Title: PARIS, an optogenetic method for functionally mapping gap junctions
    Article Snippet: Paragraph title: Molecular cloning ... The fragments were assembled using T5 exonuclease, Phusion DNA polymerase, and Taq DNA ligase (New England Biolabs).

    Fluorescence:

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Detection of doubly labeled ligation products occurs through dual fluorescence flow cytometry in the BioPlex array reader (Bio-Rad Laboratories, Hercules, CA) and leads to collection of "reporter" signal in unique allele-specific bins. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Isolation:

    Article Title: Identifying components required for OMP biogenesis as novel targets for antiinfective drugs
    Article Snippet: The vector pSB890Y and the flanking regions were then assembled using a selfmade Gibson Assembly Master Mix containing T5 Exonuclease (10U µl, Epicentre), Phusion Polymerase (2U µl, New England Biolabs) and Taq-DNA-Ligase (40U µl, New England Biolabs). .. The plasmid was then isolated using the peqGold plasmid miniprep kitI (VWR) and transformed into the diaminopimelic acid auxotroph strain E. coli ß2163.

    Article Title: Extensive sequencing of seven human genomes to characterize benchmark reference materials
    Article Snippet: Nick-labeled DNA was then incubated for 40 min at 37 °C using BioNano Genomics IrysPrep Repair mix and New England BioLabs Taq DNA Ligase at a final concentration of 1 U/μl. .. DNA was isolated from a lymphoid-cell cell culture of the Chinese son (GM24631) using the Bio-Rad CHEF Mammalian Genomic DNA Plug Kit protocol and lysed using BioNano Genomics IrysPrep Lysis Buffer and digested with QIAGEN Puregene Proteinase K. DNA was solubilized using GELase Agarose Gel-Digesting Preparation and drop-dialyzed before labeling using standard IrysPrep Reagent Kit protocols.

    Labeling:

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Detection of doubly labeled ligation products occurs through dual fluorescence flow cytometry in the BioPlex array reader (Bio-Rad Laboratories, Hercules, CA) and leads to collection of "reporter" signal in unique allele-specific bins. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Article Title: Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation
    Article Snippet: Next, the tubes were added with 1 μL of deoxyribonucleotide (25 μM) bases that are complementary with the labeling of different tubes (for example, dATP was added to tube T, and dCTP was added to tube G). .. Subsequently, 1 μL of enzyme solution (1 unit Pfu of DNA polymerase (Shanghai Sangon, Co., Ltd., China) and 2 units of Taq DNA ligase (New England BioLabs, USA) were added.

    Article Title: Integrative Detection and Analysis of Structural Variation in Cancer Genomes
    Article Snippet: 900ng DNA was digested by 30U nicking enzyme BspQ1 (New England Biolabs) in 1× buffer 3 (BioNano Genomic), 37 °C for 4 hours, and labeled with 1× labeling mix (BioNano Genomics) and 15U Taq polymerase (New England Biolabs) in 1× labeling buffer (BioNano Genomics) at 72°C for 60 minutes. .. Nick-labeled DNA was repaired in 1X repair mix (BioNano Genomics), 1× Thermo polymerase buffer (NEB), 50uM NAD+ (New England Biolabs), and 3ul 120U Taq DNA ligase (New England Biolabs) at 37°C for 30 minutes.

    Article Title: Extensive sequencing of seven human genomes to characterize benchmark reference materials
    Article Snippet: Nick-digested DNA was then incubated for 70 min at 50 °C using BioNano Genomics IrysPrep Labeling mix and New England BioLabs Taq DNA Polymerase at a final concentration of 0.4 U/μl. .. Nick-labeled DNA was then incubated for 40 min at 37 °C using BioNano Genomics IrysPrep Repair mix and New England BioLabs Taq DNA Ligase at a final concentration of 1 U/μl.

    Purification:

    Article Title: Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus
    Article Snippet: In vitro cccDNA formation assay Purified NC-DNA from Hep38.7-Tet cells was used as substrate DNA. .. NC-DNA (108 copies) was incubated with 32 units (U) of Thermostable FEN1 in ThermoPol Buffer (New England Biolabs) at 65°C for 10 min, followed by incubation with 8 U of Bst DNA polymerase, 40 U of Taq DNA ligase, 100 μM dNTPs, and NAD+ (all from New England Biolabs).

    Article Title: Integrative Detection and Analysis of Structural Variation in Cancer Genomes
    Article Snippet: Cells within plugs are lysed in 2ml cell lysis buffer (BioNano Genomics) containing 167ul proteinase K (Qiagen) for 48 hours, and washed twice with Tris-EDTA, pH 8 (TE) for 15 minutes per wash. DNA plugs were purified with 2ml 5% RNAase (Qiagen) for two hours, washed in TE for 15 minutes × 6 times, melted and equilibrated on 43°C for 45 minutes with 2ul of GELase (Epicentre). .. Nick-labeled DNA was repaired in 1X repair mix (BioNano Genomics), 1× Thermo polymerase buffer (NEB), 50uM NAD+ (New England Biolabs), and 3ul 120U Taq DNA ligase (New England Biolabs) at 37°C for 30 minutes.

    Article Title: Extensive sequencing of seven human genomes to characterize benchmark reference materials
    Article Snippet: Purified DNA in the thin layer agarose was labeled following the BioNano Genomics IrysPrep Reagent Kit protocol with adaptations for labeling in agarose. .. Nick-labeled DNA was then incubated for 40 min at 37 °C using BioNano Genomics IrysPrep Repair mix and New England BioLabs Taq DNA Ligase at a final concentration of 1 U/μl.

    Polymerase Chain Reaction:

    Article Title: Association of the MicroRNA-146a SNP rs2910164 with Ischemic Stroke Incidence and Prognosis in a Chinese Population
    Article Snippet: .. Further amplification was conducted in 10-μL multiplex LDR reaction mixtures containing 100 ng of original PCR product (1 µL), 0.05 μL of NEB Taq DNA ligase ,1 μL of probe mix, and 6.95 μL of ddH2 O, using the ABI PRISM 377 DNA Sequencer detected and GeneMapper software (ABI) analyzed. ..

    Article Title: Identifying components required for OMP biogenesis as novel targets for antiinfective drugs
    Article Snippet: Also the flanking regions ∼1000 bp up- and downstream of the target genes were amplified by PCR using the primer pairs listed in the category “Generation of deletion mutants” in Table S2. .. The vector pSB890Y and the flanking regions were then assembled using a selfmade Gibson Assembly Master Mix containing T5 Exonuclease (10U µl, Epicentre), Phusion Polymerase (2U µl, New England Biolabs) and Taq-DNA-Ligase (40U µl, New England Biolabs).

    Article Title: The Human Myotrophin Variant Attenuates MicroRNA-Let-7 Binding Ability but Not Risk of Left Ventricular Hypertrophy in Human Essential Hypertension
    Article Snippet: Reactions were performed on a thermal cycler Gene Amp PCR system 9600 (Perkin Elmer, Waltham, MA, USA). .. Further amplification was performed in a 10 μl volume of multiplex LDR reaction mixture, containing 1 μl (100 ng) of the resultant probe mix, 1 μl of probe mix, 0.05 μl of NEB Taq DNA ligase and 7.95 μl of ddH2 O.

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA). .. The multiplex LDR product (5 μL) was then added to 60 μL of hybridization solution (3 M tetramethylammonium chloride [TMAC], 50 mM Tris-HCl, pH 8.0, 3 mM EDTA, pH 8.0, 0.10% sodium dodecyl sulfate) containing 250 Luminex FlexMAP microspheres from each allelic set (total number of alleles = 9).

    Article Title: RemA is a DNA-binding protein that activates biofilm matrix gene expression in Bacillus subtilis
    Article Snippet: Insertions were verified by PCR amplification using primers 3318/3321. .. An assembly master mixture was made by combining prepared 5× isothermal assembly reaction buffer (131 mM Tris-HCl, 13.1 mM MgCl2 , 13.1 mM DTT, 8.21 mM PEG-8000, 1.32 mM NAD and 0.26 mM each dNTP) with Phusion DNA polymerase (New England BioLabs) (0.033 units µl−1 ), T5 exonuclease diluted 1:5 with 5× reaction buffer (New England BioLabs) (0.01 units µl−1 ), Taq DNA ligase (New England BioLabs) (5328 units µl−1 ) and additional dNTPs (267 µM).

    Article Title: Association of RAGE gene four single nucleotide polymorphisms with the risk, invasion, metastasis and overall survival of gastric cancer in Chinese
    Article Snippet: .. The connection reaction was performed at adding the PCR product 3 μl, 1 U 10×Taq DNA ligase buffer (New England Biolabs, United States), 0.125 μl Taq DNA ligase (40 U/μl, New England Biolabs, United States ), probe (10p) 0.01 μl/strip and with H2 O added to 10 μl under the condition of by cycles at 94℃ for 30 s, 56℃ for 3 min. LDR fluorescent products were analyzed by ABI 377 sequence analyzer (Perkin-Elmer, Foster City, CA, USA), and the results were analyzed by the fragment analysis software GeneMapper 3.7 (Applied Biosystems, Foster city, CA, USA). .. To test the accuracy of PCR-LDR method, a total of 96 DNA samples were randomly selected from the total sample, and they were genotyped using the same method, with exactly the sample findings for duplicated samples.

    Article Title: Copy number variations exploration of multiple genes in Graves’ disease
    Article Snippet: The target DNA sequences were amplified using a multiplex PCR method, and the PCR program (20 μL volume) was conducted as follows: 95°C 2 minutes, 94°C 30 s, 40 cycles (53°C 90 s, 65°C 30 s), 65°C 10 minutes. .. The ligation reaction for each sample was carried out in a final volume of 10 μL, containing 1 μL of 1× buffer, 4 μL of multi-PCR product, 1 μL of 2 pmol/μl probe mix, and 0.05 μL of 20U Taq DNA ligase (New England Biolabs).

    Article Title: Polymorphism in the Vesicular Monoamine Transporter 2 Gene Decreases the Risk of Parkinson's Disease in Han Chinese Men
    Article Snippet: .. The ligation reaction for each subject was carried out in a final volume of 20 μ L containing 20 mM Tris-HCl (pH 7.6), 25 mM potassium acetate, 10 mM magnesium acetate, 10 mM DTT, 1 mM NAD, 0.1% Triton X-100, 10 μ L of multiplex PCR product, 1 pmol of each discriminating oligo, 1 pmol of each common oligo, and 0.5 μ L of 40 U/μ L Taq DNA ligase (New England Biolabs, USA). ..

    Lysis:

    Article Title: Integrative Detection and Analysis of Structural Variation in Cancer Genomes
    Article Snippet: Cells within plugs are lysed in 2ml cell lysis buffer (BioNano Genomics) containing 167ul proteinase K (Qiagen) for 48 hours, and washed twice with Tris-EDTA, pH 8 (TE) for 15 minutes per wash. DNA plugs were purified with 2ml 5% RNAase (Qiagen) for two hours, washed in TE for 15 minutes × 6 times, melted and equilibrated on 43°C for 45 minutes with 2ul of GELase (Epicentre). .. Nick-labeled DNA was repaired in 1X repair mix (BioNano Genomics), 1× Thermo polymerase buffer (NEB), 50uM NAD+ (New England Biolabs), and 3ul 120U Taq DNA ligase (New England Biolabs) at 37°C for 30 minutes.

    Article Title: Extensive sequencing of seven human genomes to characterize benchmark reference materials
    Article Snippet: Cells were lysed using BioNano Genomics IrysPrep Lysis Buffer, protease treated with QIAGEN Puregene Proteinase K, followed by brief washing in Tris with 50 mM EDTA and then washing in Tris with 1 mM EDTA before RNase treatment with Qiagen Puregene RNase. .. Nick-labeled DNA was then incubated for 40 min at 37 °C using BioNano Genomics IrysPrep Repair mix and New England BioLabs Taq DNA Ligase at a final concentration of 1 U/μl.

    IA:

    Article Title: Biosensor libraries harness large classes of binding domains for construction of allosteric transcriptional regulators
    Article Snippet: DBDs and LBDs of this testing library were taken from the collection of DNA fragments used for the final construction of the chimeras, whereas the subset of oligonucleotides necessary to assemble them was obtained from IDT (Coralville, IA). .. This mock library enabled the testing of three different thermostable ligases: Ampligase (Epicentre, Madison, WI), 9°N DNA ligase (NEB, Ipswich, MA) and Taq DNA ligase (NEB, Ipswich, MA).

    Chromatin Immunoprecipitation:

    Article Title: Integrative Detection and Analysis of Structural Variation in Cancer Genomes
    Article Snippet: Nick-labeled DNA was repaired in 1X repair mix (BioNano Genomics), 1× Thermo polymerase buffer (NEB), 50uM NAD+ (New England Biolabs), and 3ul 120U Taq DNA ligase (New England Biolabs) at 37°C for 30 minutes. .. For each round, 160ng prepared DNA was loaded to a BioNano Irys chip that contains two flow-cells, and each round contains 30 cycles of data collection.

    Plasmid Preparation:

    Article Title: Identifying components required for OMP biogenesis as novel targets for antiinfective drugs
    Article Snippet: .. The vector pSB890Y and the flanking regions were then assembled using a selfmade Gibson Assembly Master Mix containing T5 Exonuclease (10U µl, Epicentre), Phusion Polymerase (2U µl, New England Biolabs) and Taq-DNA-Ligase (40U µl, New England Biolabs). .. Next, the strain E. coli 118λpir was transformed with the resulting plasmid.

    Software:

    Article Title: Association of the MicroRNA-146a SNP rs2910164 with Ischemic Stroke Incidence and Prognosis in a Chinese Population
    Article Snippet: .. Further amplification was conducted in 10-μL multiplex LDR reaction mixtures containing 100 ng of original PCR product (1 µL), 0.05 μL of NEB Taq DNA ligase ,1 μL of probe mix, and 6.95 μL of ddH2 O, using the ABI PRISM 377 DNA Sequencer detected and GeneMapper software (ABI) analyzed. ..

    Article Title: Association of RAGE gene four single nucleotide polymorphisms with the risk, invasion, metastasis and overall survival of gastric cancer in Chinese
    Article Snippet: .. The connection reaction was performed at adding the PCR product 3 μl, 1 U 10×Taq DNA ligase buffer (New England Biolabs, United States), 0.125 μl Taq DNA ligase (40 U/μl, New England Biolabs, United States ), probe (10p) 0.01 μl/strip and with H2 O added to 10 μl under the condition of by cycles at 94℃ for 30 s, 56℃ for 3 min. LDR fluorescent products were analyzed by ABI 377 sequence analyzer (Perkin-Elmer, Foster City, CA, USA), and the results were analyzed by the fragment analysis software GeneMapper 3.7 (Applied Biosystems, Foster city, CA, USA). .. To test the accuracy of PCR-LDR method, a total of 96 DNA samples were randomly selected from the total sample, and they were genotyped using the same method, with exactly the sample findings for duplicated samples.

    Article Title: Copy number variations exploration of multiple genes in Graves’ disease
    Article Snippet: The ligation reaction for each sample was carried out in a final volume of 10 μL, containing 1 μL of 1× buffer, 4 μL of multi-PCR product, 1 μL of 2 pmol/μl probe mix, and 0.05 μL of 20U Taq DNA ligase (New England Biolabs). .. The fluorescent products of LDR were differentiated by the software of Gene Amp PCR system 9600 (Norwalk, CT).

    Multiplex Assay:

    Article Title: Association of the MicroRNA-146a SNP rs2910164 with Ischemic Stroke Incidence and Prognosis in a Chinese Population
    Article Snippet: .. Further amplification was conducted in 10-μL multiplex LDR reaction mixtures containing 100 ng of original PCR product (1 µL), 0.05 μL of NEB Taq DNA ligase ,1 μL of probe mix, and 6.95 μL of ddH2 O, using the ABI PRISM 377 DNA Sequencer detected and GeneMapper software (ABI) analyzed. ..

    Article Title: The Human Myotrophin Variant Attenuates MicroRNA-Let-7 Binding Ability but Not Risk of Left Ventricular Hypertrophy in Human Essential Hypertension
    Article Snippet: .. Further amplification was performed in a 10 μl volume of multiplex LDR reaction mixture, containing 1 μl (100 ng) of the resultant probe mix, 1 μl of probe mix, 0.05 μl of NEB Taq DNA ligase and 7.95 μl of ddH2 O. ..

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: The 5' ends of the LDR products receive "classification" labeling in a second multiplex (Step #2) reaction where hybridization occurs between the TAG sequences added to the allele-specific primers and anti-TAG (complementary sequence) oligonucleotide probes bound to fluorescent microspheres. .. Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA).

    Article Title: Copy number variations exploration of multiple genes in Graves’ disease
    Article Snippet: The target DNA sequences were amplified using a multiplex PCR method, and the PCR program (20 μL volume) was conducted as follows: 95°C 2 minutes, 94°C 30 s, 40 cycles (53°C 90 s, 65°C 30 s), 65°C 10 minutes. .. The ligation reaction for each sample was carried out in a final volume of 10 μL, containing 1 μL of 1× buffer, 4 μL of multi-PCR product, 1 μL of 2 pmol/μl probe mix, and 0.05 μL of 20U Taq DNA ligase (New England Biolabs).

    Article Title: Polymorphism in the Vesicular Monoamine Transporter 2 Gene Decreases the Risk of Parkinson's Disease in Han Chinese Men
    Article Snippet: .. The ligation reaction for each subject was carried out in a final volume of 20 μ L containing 20 mM Tris-HCl (pH 7.6), 25 mM potassium acetate, 10 mM magnesium acetate, 10 mM DTT, 1 mM NAD, 0.1% Triton X-100, 10 μ L of multiplex PCR product, 1 pmol of each discriminating oligo, 1 pmol of each common oligo, and 0.5 μ L of 40 U/μ L Taq DNA ligase (New England Biolabs, USA). ..

    Selection:

    Article Title: The Human Myotrophin Variant Attenuates MicroRNA-Let-7 Binding Ability but Not Risk of Left Ventricular Hypertrophy in Human Essential Hypertension
    Article Snippet: Paragraph title: Gene variant selection and genotyping ... Further amplification was performed in a 10 μl volume of multiplex LDR reaction mixture, containing 1 μl (100 ng) of the resultant probe mix, 1 μl of probe mix, 0.05 μl of NEB Taq DNA ligase and 7.95 μl of ddH2 O.

    Article Title: Association of RAGE gene four single nucleotide polymorphisms with the risk, invasion, metastasis and overall survival of gastric cancer in Chinese
    Article Snippet: Paragraph title: SNP Selection and Genotyping ... The connection reaction was performed at adding the PCR product 3 μl, 1 U 10×Taq DNA ligase buffer (New England Biolabs, United States), 0.125 μl Taq DNA ligase (40 U/μl, New England Biolabs, United States ), probe (10p) 0.01 μl/strip and with H2 O added to 10 μl under the condition of by cycles at 94℃ for 30 s, 56℃ for 3 min. LDR fluorescent products were analyzed by ABI 377 sequence analyzer (Perkin-Elmer, Foster City, CA, USA), and the results were analyzed by the fragment analysis software GeneMapper 3.7 (Applied Biosystems, Foster city, CA, USA).

    In Vitro:

    Article Title: Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus
    Article Snippet: Paragraph title: In vitro cccDNA formation assay ... NC-DNA (108 copies) was incubated with 32 units (U) of Thermostable FEN1 in ThermoPol Buffer (New England Biolabs) at 65°C for 10 min, followed by incubation with 8 U of Bst DNA polymerase, 40 U of Taq DNA ligase, 100 μM dNTPs, and NAD+ (all from New England Biolabs).

    Tube Formation Assay:

    Article Title: Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus
    Article Snippet: Paragraph title: In vitro cccDNA formation assay ... NC-DNA (108 copies) was incubated with 32 units (U) of Thermostable FEN1 in ThermoPol Buffer (New England Biolabs) at 65°C for 10 min, followed by incubation with 8 U of Bst DNA polymerase, 40 U of Taq DNA ligase, 100 μM dNTPs, and NAD+ (all from New England Biolabs).

    Ethanol Precipitation:

    Article Title: Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus
    Article Snippet: NC-DNA (108 copies) was incubated with 32 units (U) of Thermostable FEN1 in ThermoPol Buffer (New England Biolabs) at 65°C for 10 min, followed by incubation with 8 U of Bst DNA polymerase, 40 U of Taq DNA ligase, 100 μM dNTPs, and NAD+ (all from New England Biolabs). .. After further incubation at 37°C for 20 min, DNA was purified by phenol/chloroform extraction and ethanol precipitation, and subjected to cccDNA-selective qPCR or RCA, as described above.

    Knock-Out:

    Article Title: Identifying components required for OMP biogenesis as novel targets for antiinfective drugs
    Article Snippet: Paragraph title: Generation of knockout strains ... The vector pSB890Y and the flanking regions were then assembled using a selfmade Gibson Assembly Master Mix containing T5 Exonuclease (10U µl, Epicentre), Phusion Polymerase (2U µl, New England Biolabs) and Taq-DNA-Ligase (40U µl, New England Biolabs).

    Concentration Assay:

    Article Title: Extensive sequencing of seven human genomes to characterize benchmark reference materials
    Article Snippet: .. Nick-labeled DNA was then incubated for 40 min at 37 °C using BioNano Genomics IrysPrep Repair mix and New England BioLabs Taq DNA Ligase at a final concentration of 1 U/μl. .. Labeled-repaired DNA was then recovered from the thin layer agarose by digesting with GELaseand counterstained with BioNano Genomics IrysPrep DNA Stain prior to data collection on the Irys system.

    Staining:

    Article Title: Integrative Detection and Analysis of Structural Variation in Cancer Genomes
    Article Snippet: Nick-labeled DNA was repaired in 1X repair mix (BioNano Genomics), 1× Thermo polymerase buffer (NEB), 50uM NAD+ (New England Biolabs), and 3ul 120U Taq DNA ligase (New England Biolabs) at 37°C for 30 minutes. .. DNA staining was finally performed with the final solution containing 1× flow buffer, 1× DTT (BioNano Genomics), and 3ul DNA stain (BioNano Genomics), in room temperature overnight.

    Article Title: Extensive sequencing of seven human genomes to characterize benchmark reference materials
    Article Snippet: Nick-labeled DNA was then incubated for 40 min at 37 °C using BioNano Genomics IrysPrep Repair mix and New England BioLabs Taq DNA Ligase at a final concentration of 1 U/μl. .. Labeled-repaired DNA was then recovered from the thin layer agarose by digesting with GELaseand counterstained with BioNano Genomics IrysPrep DNA Stain prior to data collection on the Irys system.

    Variant Assay:

    Article Title: The Human Myotrophin Variant Attenuates MicroRNA-Let-7 Binding Ability but Not Risk of Left Ventricular Hypertrophy in Human Essential Hypertension
    Article Snippet: Paragraph title: Gene variant selection and genotyping ... Further amplification was performed in a 10 μl volume of multiplex LDR reaction mixture, containing 1 μl (100 ng) of the resultant probe mix, 1 μl of probe mix, 0.05 μl of NEB Taq DNA ligase and 7.95 μl of ddH2 O.

    Luminex:

    Article Title: Differentiating Plasmodium falciparum alleles by transforming Cartesian X,Y data to polar coordinates
    Article Snippet: Individual reactions were performed in a solution (15 μL) containing 20 mM Tris-HCl buffer, pH 7.6, 25 mM potassium acetate, 10 mM magnesium acetate, 1 mM NAD+, 10 mM dithiothrietol, 0.1% Triton X-100, 10 nM (200 fmol) of each LDR probe, 1 μL of each PCR product, and 2 units of Taq DNA ligase (New England Biolabs, Beverly, MA). .. The multiplex LDR product (5 μL) was then added to 60 μL of hybridization solution (3 M tetramethylammonium chloride [TMAC], 50 mM Tris-HCl, pH 8.0, 3 mM EDTA, pH 8.0, 0.10% sodium dodecyl sulfate) containing 250 Luminex FlexMAP microspheres from each allelic set (total number of alleles = 9).

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    New England Biolabs taq dna ligase
    Enzymes and buffer components required for TEDA. ( A ) The pKat-eGFP fragment was cloned into SmaI-digested pBluescript SK–. The assembly of the two fragments was used as a model for the test. ( B ) <t>Taq</t> <t>DNA</t> ligase, Phusion DNA polymerase, T5 exonuclease (T5 exo), NAD + were tested for their necessity for the DNA assembly. In addition, Prime-STAR or FastPfu was also used instead of Phusion for testing; ( C ) PEG 8000 and dNTPs were further tested for their necessity for the DNA assembly. The concentrations of relevant components mentioned above were indicated in the figure. The base solution contained 0.1 M Tris–HCl (pH 7.5), 10 mM MgCl 2 and 10 mM dithiothreitol. The reaction was processed at 50°C for 1 h, which was the same as the Gibson assembly. *, Gibson; **, Hot Fusion; **, TEDA with dNTPs and at 50°C; ****, TEDA without dNTPs at 50°C. The data are averages of three parallel experiments with STDEV.
    Taq Dna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 96 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Enzymes and buffer components required for TEDA. ( A ) The pKat-eGFP fragment was cloned into SmaI-digested pBluescript SK–. The assembly of the two fragments was used as a model for the test. ( B ) Taq DNA ligase, Phusion DNA polymerase, T5 exonuclease (T5 exo), NAD + were tested for their necessity for the DNA assembly. In addition, Prime-STAR or FastPfu was also used instead of Phusion for testing; ( C ) PEG 8000 and dNTPs were further tested for their necessity for the DNA assembly. The concentrations of relevant components mentioned above were indicated in the figure. The base solution contained 0.1 M Tris–HCl (pH 7.5), 10 mM MgCl 2 and 10 mM dithiothreitol. The reaction was processed at 50°C for 1 h, which was the same as the Gibson assembly. *, Gibson; **, Hot Fusion; **, TEDA with dNTPs and at 50°C; ****, TEDA without dNTPs at 50°C. The data are averages of three parallel experiments with STDEV.

    Journal: Nucleic Acids Research

    Article Title: T5 exonuclease-dependent assembly offers a low-cost method for efficient cloning and site-directed mutagenesis

    doi: 10.1093/nar/gky1169

    Figure Lengend Snippet: Enzymes and buffer components required for TEDA. ( A ) The pKat-eGFP fragment was cloned into SmaI-digested pBluescript SK–. The assembly of the two fragments was used as a model for the test. ( B ) Taq DNA ligase, Phusion DNA polymerase, T5 exonuclease (T5 exo), NAD + were tested for their necessity for the DNA assembly. In addition, Prime-STAR or FastPfu was also used instead of Phusion for testing; ( C ) PEG 8000 and dNTPs were further tested for their necessity for the DNA assembly. The concentrations of relevant components mentioned above were indicated in the figure. The base solution contained 0.1 M Tris–HCl (pH 7.5), 10 mM MgCl 2 and 10 mM dithiothreitol. The reaction was processed at 50°C for 1 h, which was the same as the Gibson assembly. *, Gibson; **, Hot Fusion; **, TEDA with dNTPs and at 50°C; ****, TEDA without dNTPs at 50°C. The data are averages of three parallel experiments with STDEV.

    Article Snippet: Taq DNA ligase (New England Biolab) was used to prepare the self-produced Gibson reaction mixture.

    Techniques: Clone Assay

    Comparison of different assembly methods. ( A ) TEDA was compared with In-fusion and SLIC for the assembly of two fragments. Middle- lacZ and pBBR1MCS5::lacZ-truncated with 15-bp or 20-bp overlaps were used. 1:1, the same molar ratio of the insert to vector was used for DNA assembly; 1:2, double molar amount of the insert to vector was used for DNA assembly. ( B ) TEDA was compared with Gibson and non-optimized TEDA methods. The Pkat-eGFP and SmaI-pSK was used for cloning. TEDA(0.04U)−30°C, 0.04 U T5 exonuclease at 30°C for 40 min; TEDA(0.08 U)−30°C, 0.08 U T5 exonuclease at 30°C for 40 min; TEDA(0.04 U)−50°C, 0.04 U T5 exonuclease at 50°C for 40 min; Gibson, 0.08 U T5 exonuclease with Phusion and Taq DNA ligase at 50°C for 60 min. Neg, DNA fragments were transformed without TEDA treatment. ( C ) TEDA was compared with In-fusion for 4 fragments assembly. The 5Ptac-phbCAB operon was separated into three fragments (Figure 2A ), and they were assembled with linearized pBBR1MCS-2 to generate pBBR1MCS2::5Ptac-phbCAB. The data are averages of three parallel experiments with STDEV.

    Journal: Nucleic Acids Research

    Article Title: T5 exonuclease-dependent assembly offers a low-cost method for efficient cloning and site-directed mutagenesis

    doi: 10.1093/nar/gky1169

    Figure Lengend Snippet: Comparison of different assembly methods. ( A ) TEDA was compared with In-fusion and SLIC for the assembly of two fragments. Middle- lacZ and pBBR1MCS5::lacZ-truncated with 15-bp or 20-bp overlaps were used. 1:1, the same molar ratio of the insert to vector was used for DNA assembly; 1:2, double molar amount of the insert to vector was used for DNA assembly. ( B ) TEDA was compared with Gibson and non-optimized TEDA methods. The Pkat-eGFP and SmaI-pSK was used for cloning. TEDA(0.04U)−30°C, 0.04 U T5 exonuclease at 30°C for 40 min; TEDA(0.08 U)−30°C, 0.08 U T5 exonuclease at 30°C for 40 min; TEDA(0.04 U)−50°C, 0.04 U T5 exonuclease at 50°C for 40 min; Gibson, 0.08 U T5 exonuclease with Phusion and Taq DNA ligase at 50°C for 60 min. Neg, DNA fragments were transformed without TEDA treatment. ( C ) TEDA was compared with In-fusion for 4 fragments assembly. The 5Ptac-phbCAB operon was separated into three fragments (Figure 2A ), and they were assembled with linearized pBBR1MCS-2 to generate pBBR1MCS2::5Ptac-phbCAB. The data are averages of three parallel experiments with STDEV.

    Article Snippet: Taq DNA ligase (New England Biolab) was used to prepare the self-produced Gibson reaction mixture.

    Techniques: Plasmid Preparation, Clone Assay, Transformation Assay