New England Biolabs

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Ipswich, Massachusetts 1938

United States

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(1) Toehold-VISTA: a machine learning approach to decipher programmable RNA sensor-target interactionsNucleic Acids ResFebruary 16, 2026James M Robson, Alexander A Greentarget PCR products and a custom 2× enzyme mix containing 4 U/µl T5 exonuclease (New England Biolabs M0663S), 25 U/µl Phusion polymerase (New England Biolabs M0530S), 4 U/µl Taq ligase (New England Biolabs M0208L),

(2) Anticodon-edited transfer RNAs (ACE-tRNAs) encoded as therapeutic nonviral minimal DNA vectorsNucleic Acids ResFebruary 5, 2026Joseph J Porter, Wooree Ko, Emily G Sorensen, Zachary Cheung, Kari Thrasher, Tyler Couch, Jeffrey T Gabell, Victoria Shwe, Julia Hyatt, Jennasea B Licata, et al.room temperature. Residual unreacted MC PCR product was removed by addition of T5 exonuclease (NEB, M0663S) with ∼20 μl of 10 unit/μl T5 exonuclease added per 50 ml MC ligation reaction. The reaction containing

(3) Plasmid2MC: efficient cell-free generation of high-purity minicircle DNA for genome editing in mammalian cellsCommun BiolDecember 16, 2025Roman Teo Oliynyk, Ahmed Mahas, Emil Karpinski, George M. ChurchReagents,Reagents Component,Volume (μl) "DNA, volume X as eluted from Step 2",(X) T5 Exonuclease (NEB M0663L),4.0 "Restriction Enzyme, e.g., SspI-HF (NEB R3132L)",2.0 rCutSmart 10x buffer (NEB B6004S),10.0 Nuclease-free

(4) Anticodon Edited Transfer RNAs (ACE-tRNAs) Encoded as Therapeutic Nonviral Minimal DNA VectorsbioRxivSeptember 11, 2025Joseph J. Porter, Wooree Ko, Emily G. Sorensen, Zachary Cheung, Tyler Couch, Jeffrey T. Gabell, Victoria Shwe, Julia Hyatt, Jennasea B. Licata, Luke K. Peterson, et al.room temperature. Residual unreacted MC PCR product was removed by addition of T5 exonuclease (NEB, M0663S) with ~20 μL of 10 unit/μL T5 exonuclease added per 50 mL MC ligation reaction. The reaction containing

(5) Crosstalk between three CRISPR-Cas types enables primed type VI-A adaptation in Listeria seeligeriCell Host & MicrobeSeptember 10, 2025Shally R. Margolis, Alexander J. MeeskeM0494L Mitomycin C,VWR,CAS: 50-07-7;Cat # 22694-0020 T4 DNA Ligase,NEB,Cat # M0202L T5 exonuclease,NEB,Cat # M0663L Taq DNA ligase,NEB,Cat # M0208L Critical commercial assays,Critical commercial assays,Critical commercial

(6) Toehold-VISTA: A machine learning approach to decipher programmable RNA sensor-target interactionsbioRxivAugust 13, 2025James M. Robson, Alexander A. Greentarget PCR products and a custom 2x enzyme mix containing 4 U/μL T5 exonuclease (New England Biolabs M0663S), 25 U/μL Phusion polymerase (New England Biolabs M0530S), 4 U/μL Taq ligase (New England Biolabs M0208L),

(7) Mutations elevate an underground pathway to a physiologically relevant protopathwayMol Biol EvolAugust 7, 2025Karl A Widney, Lauren C Phillips, Leo M Rusch, Shelley D Copleyat dx.doi.org/10.17504/protocols.io.n9xdh7n using NEB Taq DNA Ligase (M0208L), NEB T5 Exonuclease (M0663L), and NEB Q5 Hi-Fi DNA Polymerase (M0491L). Five microliters of mixed fragments were added to 15 µL

(8) Development of circular AAV cargos for targeted seamless insertion with large serine integrasesMolecular Therapy Methods & Clinical DevelopmentJune 12, 2025Brett J.G. Estes, Nisha Gandhi, Jessica R. Von Stetina, Dev Paudel, Angela X. Nan, Parth Amin, Joshua Rose, Shuai Wu, Kangni Zheng, Yijun Zhang, et al.(Qiagen 129056). AAV.AD genomes were then digested by Exonuclease I (NEB M0293S) or T5 Exonuclease (NEB M0663S) and run on a native agarose gel (Invitrogen 16500500) alongside untreated genomes. For additional controls,

(9) A nucleosome switch primes hepatitis B virus infectionCellApril 17, 2025Nicholas A. Prescott, Tracy Biaco, Andrés Mansisidor, Yaron Bram, Justin Rendleman, Sarah C. Faulkner, Abigail A. Lemmon, Christine Lim, Rachel Tiersky, Eralda Salataj, et al.Nap1,"Nguyen, et al. (2022)",N/A SphI,NEB,Cat# R0182S T4 Ligase,NEB,Cat# M0202M T5 Exonuclease,NEB,Cat# M0663S TransfeX Transfection Reagent,ATCC,Cat# ACS-4005 TURBO DNase,Invitrogen,Cat# AM2238 α-amanitin,Tocris,Cat#

(10) Muskelin is a substrate adaptor of the highly regulated Drosophila embryonic CTLH E3 ligaseEMBO reportsMarch 1, 2025Briney CA, Henriksen JC, Lin C, Jones LA, Benner L, Rains AB, Gutierrez R, Gafken PR, Rissland OS.Green,Bio-RAD,1725125 Phusion,ThermoFisher Scientific,F530S Taq ligase,NEB,M0208L T5 Exonuclease,NEB,M0663S Anti-Flag ® M2 affinity gel,Millipore Sigma,A222 EZView protein G affinity beads,Millipore Sigma,E3403 Hoescht

(11) DAPE cloning with modified primers for producing designated lengths of 3' single-stranded ends in PCR productsPLoS OneFebruary 13, 2025Seoee Lee, Hyunyoung Kim, Aqsa, Kwangjin Jeung, Minho Won, Hyunju Ro(Addgene plasmid # 110403; http://n2t.net/addgene:110403; RRID:Addgene 110403). Enzymes T5 Exonuclease (M0663S), Lambda Exonuclease (M0262S), DNA Polymerase I, Large Klenow Fragment (M0210S), T4 DNA Polymerase (M0203S),

(12) Efficient sex hormone biosensors in Saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibitionSci RepJanuary 3, 2025Jie Wu, Matthias Bureik, Mario Andrea Marchisio(R0187S) restriction enzymes, O5 Hot-start polymerase (M0491S), T4 DNA ligase (M0202S), T5 exonuclease (M0663S), and Taq DNA ligase (M0208S) were purchased from New England Biolabs (NEB, USA). DNA ladder (M130-01)

(13) Living Porous Ceramics for Bacteria‐Regulated Gas Sensing and Carbon CaptureAdvanced MaterialsDecember 10, 2024Alessandro Dutto, Anton Kan, Zoubeir Saraw, Aline Maillard, Daniel Zindel, André R. Studartassembly,[ 40 ] following the protocol in reference using enzymes Taq ligase (M0208S), T5 Exonuclease (M0663S), and Phusion polymerase (M0530S), obtained from New England Biolabs (NEB). PCR fragments were obtained

(14) Control of DNA replication in vitro using a reversible replication barrierNature ProtocolsJuly 1, 2024Emma J. Vontalge, Tamar Kavlashvili, Steven N. Dahmen, Matthew T. Cranford, James M. Dewarr3.1 (10X) (NEB, cat. no. B7203S) AgeI-HF (NEB, cat. no. R3552S) T5 Exonuclease (NEB, cat. no. M0663S) Ethidium bromide (Invitrogen, cat. no. 15585011) CAUTION Ethidium bromide is a mutagen. Wear proper

(15) Brain-wide silencing of prion protein by AAV-mediated delivery of an engineered compact epigenetic editorScienceJune 28, 2024Edwin N. Neumann, Tessa M. Bertozzi, Elaine Wu, Fiona Serack, John W. Harvey, Pamela P. Brauer, Catherine P. Pirtle, Alissa Coffey, Michael Howard, Nikita Kamath, et al.Trizol-Chloroform extraction from brain homogenate followed by treatment with T5 exonuclease (New England BioLabs, M0663S) and RNase Cocktail Enzyme Mix (Thermo Fisher Scientific, AM2288). To extract single-stranded AAV DNA,

(16) Ribosome profiling reveals the role of yeast RNA-binding proteins Cth1 and Cth2 in translational regulationiScienceJune 21, 2024Barlit H, Romero AM, Gülhan A, Patnaik PK, Tyshkovskiy A, Martínez-Pastor MT, Gladyshev VN, Puig S, Labunskyy VM.(PEG-8000)",ThermoFisher,Cat#043443.36 Taq DNA Ligase,New England Biolabs,Cat#M0208S T5 exonuclease,New England Biolabs,Cat#M0663S BsaI-HFv2,New England Biolabs,Cat#R3733S ,, Deposited data,Deposited data,Deposited data ,, Raw

(17) Manipulating mRNA-binding protein Cth2 function in budding yeast Saccharomyces cerevisiaeSTAR protocolsMarch 15, 2024Patnaik PK, Barlit H, Labunskyy VM.,Thermo Fisher Scientific,Cat#043443.36 Taq DNA ligase,New England Biolabs,Cat#M0208S T5 exonuclease,New England Biolabs,Cat#M0663S BsaI-HFv2,New England Biolabs,Cat#R3733S ,, Experimental models: Organisms/strains,Experimental models:

(18) High-fidelity 3D live-cell nanoscopy through data-driven enhanced super-resolution radial fluctuationNature MethodsNovember 13, 2023Romain F. Laine, Hannah S. Heil, Simao Coelho, Jonathon Nixon-Abell, Angélique Jimenez, Theresa Wiesner, Damián Martínez, Tommaso Galgani, Louise Régnier, Aki Stubb, et al.#8504574) to Skylan-NS (Addgene plasmid #8678558) by Gibson assembly75 with a 0.005 U µl−1 T5 exonuclease (M0663S, NEB), 0.033 U µl−1 Phusion DNA polymerase (F-553L, Finnzymes) and 5.33 U µl−1 TAQ DNA ligase (MB42601,

(19) Targeted viral adaptation generates a simian-tropic hepatitis B virus that infects marmoset cellsNature CommunicationsJune 16, 2023Yongzhen Liu, Thomas R. Cafiero, Debby Park, Abhishek Biswas, Benjamin Y. Winer, Cheul H. Cho, Yaron Bram, Vasuretha Chandar, Aoife K. O’ Connell, Hans P. Gertje, et al.the samples from 2 wells for each group were combined. Then 5 μL Hirt DNA was used for T5 treatment (#M0663S, NEB, Ipswich, MA) for 30 min at 37 °C in a 10 μL reaction system with 0.5 μL T5 exonuclease followed

(20) Structural basis for the assembly of the type V CRISPR-associated transposon complexCellDecember 22, 2022Michael Schmitz, Irma Querques, Seraina Oberli, Christelle Chanez, Martin JinekdATP solution,Roth,Cat#K035.1 dTTP solution,Roth,Cat#K036.1 NAD,Roth,Cat#AE11.1 T5 exonuclease,New England Biolabs,Cat#M0663S Phusion® High-Fidelity DNA Polymerase,New England Biolabs,Cat#M0530L Taq DNA Ligase,New England Biolabs,Cat#M0208L T4 DNA Ligase

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T5 Exonuclease (Category 4)

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T5 Exonuclease (Category 4) from New England Biolabs

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