large klenow fragment (New England Biolabs)

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New England Biolabs large klenow fragment
Large Klenow Fragment, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1626 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/large+klenow+fragment/DNA+Pol+I+Klenow+Fragment/pm42827183-274-12-15
Average 99 stars, based on 1626 article reviews

large klenow fragment - by Bioz Stars, 2026-11

99/100 stars

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Labeling:

Article Title: 3D Chromatin Architecture Provides Insights Into Leaf Trait Variation Among Pear Species.
Article Snippet: Subsequently, the cross-linked nuclei were permeated with 0.3% SDS at 65◦C for 10 min. SDS were quenched by 1.8% Triton X100 at 37◦C for 15 min. Chromatin was then digested with 100 U of MboI (NEB) restriction enzyme for 2 h at 37◦C. .. DNA ends were labeled with biotin-14-dCTP (Invitrogen) by incubating with DNA Polymerase I, Large Klenow Fragment (NEB) at 37◦C for 45 min, after which the reaction was inactivated by incubation at 65◦C for 20 min. DNA ligation was performed by 50U T4 DNA ligase (Thermo) at 16◦C for 4 h. Afterward, proteinase K was added to the sample and incubated at 65◦C overnight to reverse the cross-linking. .. DNA was extracted using the Qiagen DNeasy Plant Mini Kit (Qiagen, #69106), following the manufacturer’s instructions.

Article Title: Cooperative Roles of Pds5a and Pds5b Constrain Long‐Range Chromatin Interactions in Vertebrate Embryos
Article Snippet: The lysate was centrifuged at 2500 g for 5 min, washed once with ice‐cold Hi‐C lysis buffer, resuspended in 50 μL of 0.5% SDS, and incubated at 62°C for 10 min. After adding 145 μL of water and 25 μL of 10% Triton X‐100, samples were incubated at 37°C for 15 min. Next, 25 μL of 10× NEBuffer 2 and 100 U of MboI were added, and chromatin was digested at 37°C for 2 h with rotation. .. MboI was inactivated by incubation at 62°C for 20 min. DNA ends were labeled with biotin by adding 50 μL of fill‐in master mix (37.5 μL of 0.4 mM biotin‐14‐dATP [Life Technologies], 1.5 μL each of 10 mM dCTP, dGTP, and dTTP, and 8 μL of 5 U/μL DNA Polymerase I Large Klenow Fragment [NEB]) and incubating at 37°C for 1.5 h with rotation. ..

Incubation:

Article Title: Unraveling the origin of the Azorean viral nervous necrosis 2024 outbreak through spatiotemporal and time-resolved phylogenetic analysis
Article Snippet: The final mixture was incubated at 23°C for 10 min, followed by 50°C for 45 min and 80°C for 10 min, in a T100 Thermal Cycler (Bio-Rad Laboratories, USA). .. To eliminate residual RNA molecules, 1 mL of E. coli RNase H (2 U/mL) was added to the mixture, which was then incubated at 37 °C for 20 min. To complete the formation of double-stranded cDNA molecules, 1 mL (5 U) of DNA Polymerase I Large (Klenow) Fragment (New England Biolabs Inc., USA), 2.5 mL of 10X NEBuffer 2 (New England Biolabs Inc., Ipswich, USA), and 0.5 mL of dNTP mix (10 mM) were added to 20 mL of cDNA. ..

Article Title: Chromatin interaction landscape of epimorphic regeneration in teleost fish
Article Snippet: To enhance the efficiency of T4 DNA ligation, the cell pellet was incubated with 50 μl of premix solution [5 μl of 10× NEBuffer 2.1, 1 μl of 100 mM adenosine triphosphate (ATP), 2.5 μl of 100 mM dithiothreitol, 34 μl of H 2 O, and 2.5 μl of T4 PNK (10 U/μl)] for 15 min at 37°C. .. Then, large Klenow fragment (NEB, M0210L) was added with incubation for another 15 min. After that, biotinylated nucleotides were incorporated by incubating the sample with 25 μl of premix buffer [5 μl of 1 mM biotin-dATP, 5 μl of 1 mM biotin–2′-deoxycytidine 5′-triphosphate, 0.5 μl of 10 mM dTTP + 2′-deoxyguanosine 5′-triphosphate, 2.5 μl of 10× T4 DNA ligase buffer, 0.125 μl of bovine serum albumin (BSA; 20 mg/ml), and 11.875 μl of ddH 2 O] for 45 min at RT. ..

Article Title: Single-allele nanoscale mapping of regulatory variants.
Article Snippet: The remaining reaction was centrifuged, PBS washed and resuspended in DNA ligation buffer (Thermo Fisher Scientific; final concentrations: 40 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 10 mM dithiothreitol and 5 mM ATP) supplemented with dNTPs (final concentration 400 μM each for dATP, dCTP, dGTP and dTTP (Thermo Fisher Scientific, cat. no. R0191)) and EGTA 5 mM. .. T4 polynucleotide kinase PNK (NEB, cat. no. M0201L) and DNA polymerase I large (Klenow) fragment (NEB, cat. no. M0210L) were at final concentrations of 200 U ml−1 and 100 U ml−1, respectively, T4 DNA ligase (Thermo Fisher Scientific, high-concentration ligase (30 U μl−1), cat. no. EL0013) to a final concentration of 300 U ml−1 was added and the reaction was incubated at 37 °C for 2 h before cooling to 20 °C and overnight incubation on an Eppendorf Thermomixer with intermittent 500-r.p.m. ..

DNA Ligation:

other:

Article Title: A critical role of the histone lysine demethylase JmjC1 in regulating severe malaria-associated virulence genes in Plasmodium falciparum.
Article Snippet: Cross-linked DNA was digested with MboI (NEB), and 5’ overhangs were filled in using dNTPs supplementedwith Biotin-14-dCTP (Invitrogen) andDNAPolymerase I, Large (Klenow) Fragment (NEB).

Polymerase Chain Reaction:

Article Title: Pooled CRISPRi screening reveals fungal-specific drug target candidates.
Article Snippet: The rising rate of drug-resistant fungal infections and the emergence of intrinsically resistant pathogens pose growing clinical challenges.. Because fungi are closely related to mammals, developing antifungals without toxic off-target effects is difficult.. Targeted gene repression can model drug-mediated inhibition and reveal gene dosage sensitivity, but traditional approaches in the fungal pathogen Candida albicans are labour intensive and low throughput.

Concentration Assay:

Purification:

Article Title: Influenza A virus NS1 sequesters RNA:DNA hybrids to evade RNase H1-dependent innate immunity
Article Snippet: .. Second-strand synthesis was performed on ssDNA using random hexamers, dNTPs, and the Large (Klenow) Fragment (NEB) according to the manufacturer’s instructions, followed by purification with 2x volume AMPure XP beads (Beckman Coulter). .. Sequencing libraries were prepared from the resulting dsDNA using the Nextera XT DNA Library Preparation Kit and sequenced on the MiSeq platform (Illumina, to a depth of ≥1 5 single-end 151-bp reads).

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