klenow fragment  (New England Biolabs)


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    Structured Review

    New England Biolabs klenow fragment
    Klenow Fragment, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/klenow fragment/product/New England Biolabs
    Average 99 stars, based on 79 article reviews
    Price from $9.99 to $1999.99
    klenow fragment - by Bioz Stars, 2020-02
    99/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Arginine starvation kills tumor cells through aspartate exhaustion and mitochondrial dysfunction
    Article Snippet: dNTP measurement Cells (1 × 106 ) were extracted with freon/trioctylamine (55%:45%) and vortexed, followed by centrifugation at 20,000×g for 5 min. .. The reaction mixture (100 μL) contained 40 mM Tris-HCl (pH 7.4), MgCl2 (10 mM), dithiothereitol (5 mM), oligonucleotide template (0.25 μM), RNase A (1.5 μg), 3 H-dATP (for dTTP, dCTP, and dGTP) or 3 H-dTTP (for dATP) (0.25 μM), Klenow fragment (0.2 units, NEB, for dTTP), Klenow fragment (0.025 units, NEB, for dATP), or Taq polymerase (2 units, Zgene, for dCTP and dGTP) and 16 μL cell extract.

    Synthesized:

    Article Title: Genome-wide mapping of alternative splicing in Arabidopsis thaliana
    Article Snippet: .. The second strand of cDNA was synthesized using DNA polymerase I (Klenow fragment) by combining 20 μL of the first strand reaction, 8 μL of 10× Klenow Buffer (NEB), 1 unit of RNase H (Invitrogen), 68.8 μL of water, and 30 units of DNA polymerase I (NEB). .. The reaction was incubated for 90 min at 15°C and cDNA was purified using the QIAquick PCR clean up kit.

    Ethanol Precipitation:

    Article Title: A method for multi-codon scanning mutagenesis of proteins based on asymmetric transposons
    Article Snippet: The digestion product of each library was treated with Klenow fragment by adding 1.7 μl of 1 mM dNTPs and 0.5 U of Klenow fragment (NEB) into the heat-inactivated reaction solution. .. The ligation reaction was incubated at 16°C overnight and then heat inactivated at 70°C for 7 min. Several 10 μl ligation reactions were performed for each library, pooled and then purified by ethanol precipitation.

    Ligation:

    Article Title: A method for multi-codon scanning mutagenesis of proteins based on asymmetric transposons
    Article Snippet: The digestion product of each library was treated with Klenow fragment by adding 1.7 μl of 1 mM dNTPs and 0.5 U of Klenow fragment (NEB) into the heat-inactivated reaction solution. .. The Klenow-treated digestion product was gel-purified using QIAquick gel-extraction kit (Qiagen) and then subjected to an intra-molecular ligation that contained 2.5 ng/μl DNA, 0.5 Weiss unit/μl T4 DNA ligase (Fermentas), 40 mM Tris-HCl (pH 7.8 at 25°C), 10 mM MgCl2 , 10 mM DTT and 0.5 mM ATP in a 10 μl reaction.

    Incubation:

    Article Title: Arginine starvation kills tumor cells through aspartate exhaustion and mitochondrial dysfunction
    Article Snippet: The reaction mixture (100 μL) contained 40 mM Tris-HCl (pH 7.4), MgCl2 (10 mM), dithiothereitol (5 mM), oligonucleotide template (0.25 μM), RNase A (1.5 μg), 3 H-dATP (for dTTP, dCTP, and dGTP) or 3 H-dTTP (for dATP) (0.25 μM), Klenow fragment (0.2 units, NEB, for dTTP), Klenow fragment (0.025 units, NEB, for dATP), or Taq polymerase (2 units, Zgene, for dCTP and dGTP) and 16 μL cell extract. .. Incubation was carried out for 60 min at 37 °C for dTTP and dATP, or at 48 °C for dCTP and dGTP, and reaction mixtures were spotted onto Zeta-probe blotting membrane (Bio-Rad).

    Article Title: A method for multi-codon scanning mutagenesis of proteins based on asymmetric transposons
    Article Snippet: The reaction was incubated at 37°C for 4.5 h followed by heat inactivation at 65°C for 20 min. .. The digestion product of each library was treated with Klenow fragment by adding 1.7 μl of 1 mM dNTPs and 0.5 U of Klenow fragment (NEB) into the heat-inactivated reaction solution.

    Article Title: Genome-wide mapping of alternative splicing in Arabidopsis thaliana
    Article Snippet: The second strand of cDNA was synthesized using DNA polymerase I (Klenow fragment) by combining 20 μL of the first strand reaction, 8 μL of 10× Klenow Buffer (NEB), 1 unit of RNase H (Invitrogen), 68.8 μL of water, and 30 units of DNA polymerase I (NEB). .. The reaction was incubated for 90 min at 15°C and cDNA was purified using the QIAquick PCR clean up kit.

    Purification:

    Article Title: A method for multi-codon scanning mutagenesis of proteins based on asymmetric transposons
    Article Snippet: The purified PCR product of di-codon or tri-codon library was digested with BsgI in a 50 μl reaction mixture containing 500 ng DNA, 6 U BsgI (NEB), 80 μM S -adenosylmethionine (SAM) and 1× buffer 4 (NEB). .. The digestion product of each library was treated with Klenow fragment by adding 1.7 μl of 1 mM dNTPs and 0.5 U of Klenow fragment (NEB) into the heat-inactivated reaction solution.

    Article Title: Genome-wide mapping of alternative splicing in Arabidopsis thaliana
    Article Snippet: The second strand of cDNA was synthesized using DNA polymerase I (Klenow fragment) by combining 20 μL of the first strand reaction, 8 μL of 10× Klenow Buffer (NEB), 1 unit of RNase H (Invitrogen), 68.8 μL of water, and 30 units of DNA polymerase I (NEB). .. The reaction was incubated for 90 min at 15°C and cDNA was purified using the QIAquick PCR clean up kit.

    Random Hexamer Labeling:

    Article Title: Genome-wide mapping of alternative splicing in Arabidopsis thaliana
    Article Snippet: For the RP cDNA libraries, the first cDNA strand was synthesized using 1 μg of poly(A) mRNA essentially free of rRNA, random hexamer primers (300 ng per microgram of RNA), and Superscript III reverse transcriptase (RT) (Invitrogen). .. The second strand of cDNA was synthesized using DNA polymerase I (Klenow fragment) by combining 20 μL of the first strand reaction, 8 μL of 10× Klenow Buffer (NEB), 1 unit of RNase H (Invitrogen), 68.8 μL of water, and 30 units of DNA polymerase I (NEB).

    Polymerase Chain Reaction:

    Article Title: A method for multi-codon scanning mutagenesis of proteins based on asymmetric transposons
    Article Snippet: The purified PCR product of di-codon or tri-codon library was digested with BsgI in a 50 μl reaction mixture containing 500 ng DNA, 6 U BsgI (NEB), 80 μM S -adenosylmethionine (SAM) and 1× buffer 4 (NEB). .. The digestion product of each library was treated with Klenow fragment by adding 1.7 μl of 1 mM dNTPs and 0.5 U of Klenow fragment (NEB) into the heat-inactivated reaction solution.

    Article Title: Genome-wide mapping of alternative splicing in Arabidopsis thaliana
    Article Snippet: The second strand of cDNA was synthesized using DNA polymerase I (Klenow fragment) by combining 20 μL of the first strand reaction, 8 μL of 10× Klenow Buffer (NEB), 1 unit of RNase H (Invitrogen), 68.8 μL of water, and 30 units of DNA polymerase I (NEB). .. The reaction was incubated for 90 min at 15°C and cDNA was purified using the QIAquick PCR clean up kit.

    Gel Extraction:

    Article Title: A method for multi-codon scanning mutagenesis of proteins based on asymmetric transposons
    Article Snippet: The digestion product of each library was treated with Klenow fragment by adding 1.7 μl of 1 mM dNTPs and 0.5 U of Klenow fragment (NEB) into the heat-inactivated reaction solution. .. The Klenow-treated digestion product was gel-purified using QIAquick gel-extraction kit (Qiagen) and then subjected to an intra-molecular ligation that contained 2.5 ng/μl DNA, 0.5 Weiss unit/μl T4 DNA ligase (Fermentas), 40 mM Tris-HCl (pH 7.8 at 25°C), 10 mM MgCl2 , 10 mM DTT and 0.5 mM ATP in a 10 μl reaction.

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    New England Biolabs restriction endonucleases mse i
    Restriction profiles obtained after digestion of polymerase chain reaction products from the ITS1 locus with <t>Mse</t> I. lanes 1- 3-, 100-, 50-, and 10-bp ladder molecular weight markers; lanes 4 and 15, bovine genotype 2 isolates; lanes 5, 6, 14, and 16, human genotype 1 isolates including DE340 (lane 14); lanes 7 and 9–12, cervine genotype isolates including MH205 (lane 7), TK320 (lane 10), and DE302 (lane 11); lane 8, Cryptosporidium meleagridis isolate CS33; lanes 13 and 17, other novel genotype isolates such as VF383 (lane 13) and TK348 (lane 17)
    Restriction Endonucleases Mse I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 76/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    New England Biolabs restriction endonuclease fsp i
    Agarose gel electrophoresis of KS7-KS8 <t>PCR</t> products digested with <t>Fsp</t> I (A) or Hha I (B) and of PCR amplification products with primers Stx1c-1 and Stx1c-2 (C). M, molecular weight marker (1-kb DNA ladder; Gibco BRL, Eggenstein, Germany). In lanes 1 to 12, the following STEC strains (genotypes, serotypes, and origins, if not human, in parentheses) are shown: 1, 808/97 ( stx 1c + stx 2d ; ONT:H8); 2, 3115/97 ( stx 1c + stx 2d ; O128:H2); 3, 521/99 ( stx 1c + stx 2d ; O rough :H19); 4, 4756/98 ( stx 1c + stx 2d ; O70:H − ); 5, 273/00 ( stx 1c + stx 2d ; O128:H − ; sheep); 6, 295/00 ( stx 1c + stx 2d ; O128:H − ; sheep); 7, EDL 933 ( stx 1 + stx 2 ; O157:H7); 8, 2544/00 ( stx 1 ; O145:H − ); 9, 3385/00 ( stx 1 + stx 2 ; O111:H − ); 10, 4424/99 ( stx 1 + stx 2 ; O157:H7); 11, 2049/98 ( stx 1 + stx 2c ; O157:H − ); 12, 2050/98 ( stx 1 + stx 2 + stx 2c ; O157:H − ).
    Restriction Endonuclease Fsp I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction endonuclease fsp i/product/New England Biolabs
    Average 78 stars, based on 1 article reviews
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    78
    New England Biolabs restriction endonuclease bstz17 i
    A – Non-radioactive PCR-SSCP analysis of VHL exon 3; T – altered mobility observed in a tumor sample, relative to the normal control N. B – Ethidium bromide-stained polyacrylamide gel of <t>BstZ17</t> I restriction digestion. Lane 1 – peripheral venous blood, Lane 2 – renal cell carcinoma, Lane 3 – non tumoral tissue from kidney, Lane 4 – thyroid metastases, M – denotes the lane containing the pUC Mix Marker 8 (Fermentas, Burlingyon, Canada). For the wild allele, restriction digestion produces bands of 212 and 59 bp. The mutation abolishes the restriction site and the mutant allele corresponds to a 271 bp band. C and D – Tumour DNA sequence in the VHL codon 156 region of cloned exon 3 amplicons. As a consequence of the 680 delA, a premature stop codon appears in the mutant allele.
    Restriction Endonuclease Bstz17 I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction endonuclease bstz17 i/product/New England Biolabs
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    New England Biolabs restriction endonuclease bsrd i
    Restriction fragment length polymorphism patterns obtained upon DNA digestion with <t>BsrD</t> I. M: 100-base pair (bp) marker, 1: Microsporumcanis (IFM45829), 2: M. gypseum (IFM-5292), 3: Trichophyton mentagrophytes var. interdigitale (IFM-48155), 4: T. mentagrophytes var. mentagrophytes (CBS113880), 5: T. rubrum (ATCC28188), 6: T. tonsurans (CBS109036).
    Restriction Endonuclease Bsrd I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Restriction profiles obtained after digestion of polymerase chain reaction products from the ITS1 locus with Mse I. lanes 1- 3-, 100-, 50-, and 10-bp ladder molecular weight markers; lanes 4 and 15, bovine genotype 2 isolates; lanes 5, 6, 14, and 16, human genotype 1 isolates including DE340 (lane 14); lanes 7 and 9–12, cervine genotype isolates including MH205 (lane 7), TK320 (lane 10), and DE302 (lane 11); lane 8, Cryptosporidium meleagridis isolate CS33; lanes 13 and 17, other novel genotype isolates such as VF383 (lane 13) and TK348 (lane 17)

    Journal: Emerging Infectious Diseases

    Article Title: Novel Cryptosporidium Genotypes in Sporadic Cryptosporidiosis Cases: First Report of Human Infections with a Cervine Genotype

    doi: 10.3201/eid0803.010194

    Figure Lengend Snippet: Restriction profiles obtained after digestion of polymerase chain reaction products from the ITS1 locus with Mse I. lanes 1- 3-, 100-, 50-, and 10-bp ladder molecular weight markers; lanes 4 and 15, bovine genotype 2 isolates; lanes 5, 6, 14, and 16, human genotype 1 isolates including DE340 (lane 14); lanes 7 and 9–12, cervine genotype isolates including MH205 (lane 7), TK320 (lane 10), and DE302 (lane 11); lane 8, Cryptosporidium meleagridis isolate CS33; lanes 13 and 17, other novel genotype isolates such as VF383 (lane 13) and TK348 (lane 17)

    Article Snippet: RFLP Analyses of PCR Products PCR products were purified by using QIAquick spin columns (Qiagen, Mississauga, ON) according to the manufacturer’s instructions before digestion with the restriction endonucleases Mse I (New England BioLabs, Mississauga, ON) for the ITS1 locus and Rsa I (New England BioLabs) for the COWP gene.

    Techniques: Polymerase Chain Reaction, Molecular Weight

    Agarose gel electrophoresis of KS7-KS8 PCR products digested with Fsp I (A) or Hha I (B) and of PCR amplification products with primers Stx1c-1 and Stx1c-2 (C). M, molecular weight marker (1-kb DNA ladder; Gibco BRL, Eggenstein, Germany). In lanes 1 to 12, the following STEC strains (genotypes, serotypes, and origins, if not human, in parentheses) are shown: 1, 808/97 ( stx 1c + stx 2d ; ONT:H8); 2, 3115/97 ( stx 1c + stx 2d ; O128:H2); 3, 521/99 ( stx 1c + stx 2d ; O rough :H19); 4, 4756/98 ( stx 1c + stx 2d ; O70:H − ); 5, 273/00 ( stx 1c + stx 2d ; O128:H − ; sheep); 6, 295/00 ( stx 1c + stx 2d ; O128:H − ; sheep); 7, EDL 933 ( stx 1 + stx 2 ; O157:H7); 8, 2544/00 ( stx 1 ; O145:H − ); 9, 3385/00 ( stx 1 + stx 2 ; O111:H − ); 10, 4424/99 ( stx 1 + stx 2 ; O157:H7); 11, 2049/98 ( stx 1 + stx 2c ; O157:H − ); 12, 2050/98 ( stx 1 + stx 2 + stx 2c ; O157:H − ).

    Journal: Journal of Clinical Microbiology

    Article Title: Identification, Characterization, and Distribution of a Shiga Toxin 1 Gene Variant (stx1c) in Escherichia coli Strains Isolated from Humans

    doi: 10.1128/JCM.40.4.1441-1446.2002

    Figure Lengend Snippet: Agarose gel electrophoresis of KS7-KS8 PCR products digested with Fsp I (A) or Hha I (B) and of PCR amplification products with primers Stx1c-1 and Stx1c-2 (C). M, molecular weight marker (1-kb DNA ladder; Gibco BRL, Eggenstein, Germany). In lanes 1 to 12, the following STEC strains (genotypes, serotypes, and origins, if not human, in parentheses) are shown: 1, 808/97 ( stx 1c + stx 2d ; ONT:H8); 2, 3115/97 ( stx 1c + stx 2d ; O128:H2); 3, 521/99 ( stx 1c + stx 2d ; O rough :H19); 4, 4756/98 ( stx 1c + stx 2d ; O70:H − ); 5, 273/00 ( stx 1c + stx 2d ; O128:H − ; sheep); 6, 295/00 ( stx 1c + stx 2d ; O128:H − ; sheep); 7, EDL 933 ( stx 1 + stx 2 ; O157:H7); 8, 2544/00 ( stx 1 ; O145:H − ); 9, 3385/00 ( stx 1 + stx 2 ; O111:H − ); 10, 4424/99 ( stx 1 + stx 2 ; O157:H7); 11, 2049/98 ( stx 1 + stx 2c ; O157:H − ); 12, 2050/98 ( stx 1 + stx 2 + stx 2c ; O157:H − ).

    Article Snippet: The stxB 1 gene was amplified with primers KS7-KS8 (Table ), and 12 μl of each PCR product was digested with restriction endonuclease Fsp I or Hha I (New England BioLabs GmbH, Frankfurt, Germany), as recommended by the manufacturer.

    Techniques: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Amplification, Molecular Weight, Marker

    A – Non-radioactive PCR-SSCP analysis of VHL exon 3; T – altered mobility observed in a tumor sample, relative to the normal control N. B – Ethidium bromide-stained polyacrylamide gel of BstZ17 I restriction digestion. Lane 1 – peripheral venous blood, Lane 2 – renal cell carcinoma, Lane 3 – non tumoral tissue from kidney, Lane 4 – thyroid metastases, M – denotes the lane containing the pUC Mix Marker 8 (Fermentas, Burlingyon, Canada). For the wild allele, restriction digestion produces bands of 212 and 59 bp. The mutation abolishes the restriction site and the mutant allele corresponds to a 271 bp band. C and D – Tumour DNA sequence in the VHL codon 156 region of cloned exon 3 amplicons. As a consequence of the 680 delA, a premature stop codon appears in the mutant allele.

    Journal: BMC Endocrine Disorders

    Article Title: A multinodular goiter as the initial presentation of a renal cell carcinoma harbouring a novel VHL mutation

    doi: 10.1186/1472-6823-6-6

    Figure Lengend Snippet: A – Non-radioactive PCR-SSCP analysis of VHL exon 3; T – altered mobility observed in a tumor sample, relative to the normal control N. B – Ethidium bromide-stained polyacrylamide gel of BstZ17 I restriction digestion. Lane 1 – peripheral venous blood, Lane 2 – renal cell carcinoma, Lane 3 – non tumoral tissue from kidney, Lane 4 – thyroid metastases, M – denotes the lane containing the pUC Mix Marker 8 (Fermentas, Burlingyon, Canada). For the wild allele, restriction digestion produces bands of 212 and 59 bp. The mutation abolishes the restriction site and the mutant allele corresponds to a 271 bp band. C and D – Tumour DNA sequence in the VHL codon 156 region of cloned exon 3 amplicons. As a consequence of the 680 delA, a premature stop codon appears in the mutant allele.

    Article Snippet: Restriction analysis, using the restriction endonuclease BstZ17 I (New England BioLabs® , Inc., Beverly, USA) was also performed.

    Techniques: Polymerase Chain Reaction, Staining, Marker, Mutagenesis, Sequencing, Clone Assay

    Restriction fragment length polymorphism patterns obtained upon DNA digestion with BsrD I. M: 100-base pair (bp) marker, 1: Microsporumcanis (IFM45829), 2: M. gypseum (IFM-5292), 3: Trichophyton mentagrophytes var. interdigitale (IFM-48155), 4: T. mentagrophytes var. mentagrophytes (CBS113880), 5: T. rubrum (ATCC28188), 6: T. tonsurans (CBS109036).

    Journal: Annals of Dermatology

    Article Title: Identification of Dermatophytes by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Analysis of Metalloproteinase-1

    doi: 10.5021/ad.2014.26.3.338

    Figure Lengend Snippet: Restriction fragment length polymorphism patterns obtained upon DNA digestion with BsrD I. M: 100-base pair (bp) marker, 1: Microsporumcanis (IFM45829), 2: M. gypseum (IFM-5292), 3: Trichophyton mentagrophytes var. interdigitale (IFM-48155), 4: T. mentagrophytes var. mentagrophytes (CBS113880), 5: T. rubrum (ATCC28188), 6: T. tonsurans (CBS109036).

    Article Snippet: 5) Detection of restriction fragment length polymorphism in the metalloproteinase-1 regions Ten micrograms of purified PCR sample was digested at 65℃ for 16 hours with 10 U of restriction endonuclease BsrD I (New England Biolabs, Ipswich, MA, USA).

    Techniques: Marker