Enzymes and Inhibitors
Enzymes and Inhibitors
New England Biolabs, Inc. StickTogether™ DNA Ligase Buffer – 2 ml
New England Biolabs supplies a reaction buffer with all of its enzymes. At a 1X concentration this reaction buffer assures optimal activity of T3, T7, Hi-T4 and Salt-T4 DNA Ligases .
New England Biolabs, Inc. Exonuclease I Reaction Buffer – 6 ml
This is the NEB reaction buffer supplied with Exonuclease I. New England Biolabs supplies a 10X reaction buffer with all of its enzymes. At a 1X concentration this reaction buffer assures optimal activity of the enzyme.
New England Biolabs, Inc. rCutSmart™ Buffer - 5 mls
New England Biolabs provides a color-coded 10X NEBuffer with each restriction endonuclease to ensure optimal (100%) activity. Most of our enzymes are supplied with one of four standard NEBuffers. These buffers contain Recombinant Albumin (NEB #B9200). Over 215 restriction enzymes are 100% active in rCutSmart Buffer, making it significantly easier to set up your double digest reactions. Since rCutSmart Buffer includes Recombinant Albumin, there are also fewer tubes and pipetting steps to worry about. Additionally, many DNA modifying enzymes are 100% active in rCutSmart Buffer, eliminating the need for subsequent purification.
New England Biolabs, Inc. PaqCI - 200 units
PaqCI is a Type IIS restriction enzyme with a 7 basepair recognition sequence: CACCTGC(4/8). It is sourced from an E. coli strain that carries the modified cloned PaqCI gene from Paucibacter aquatile. PaqCI requires multiple sites for efficient cleavage -- PaqCI Activator is included for optimal cleavage. This enzyme is supplied with NEB's rCutSmart Buffer and is heat inactivated.
New England Biolabs, Inc. PluTI – 500 units
A restriction endonuclease that recognizes the sequence GGCGC^C
New England Biolabs, Inc. Thermostable USER III Enzyme - 50 units
Thermostable USER (Uracil-Specific Excision Reagent) III Enzyme generates a single nucleotide gap at the location of a uracil. Thermostable USER III Enzyme is a mixture of Afu Uracil DNA glycosylase (UDG) and the DNA glycosylase-lyase Endonuclease IV. Afu UDG catalyzes the excision of a uracil base, forming an abasic (apyrimidinic) site while leaving the phosphodiester backbone intact. The lyase activity of Endonuclease IV breaks the phosphodiester backbone at the 3’ and 5’ sides of the abasic site so that base-free deoxyribose is released leaving a 5’ deoxyribose phosphate (dRP) and a 3’-hydroxyl. Thermostable USER III Enzyme also has 3’-diesterase activity which can remove 3’ phosphate, 3’-alpha, beta-unsaturated aldehyde, phosphoglycoaldehyde, and other 3' blocking groups. Thermostable USER III Enzyme is optimally active at 65C.
New England Biolabs, Inc. EnGen Spy Cas9 HF1 - 500 pmol
EnGen Spy Cas9 HF1 is a high-fidelity, quadruple substitution (N497A/R661A/Q695A/Q926A) variant of EnGen Spy Cas9 NLS from Streptococcus pyogenes with reduced non-specific DNA cleavage. Spy Cas9 is an RNA-guided endonuclease that catalyzes site-specific cleavage of double stranded DNA. The single guide RNA (sgRNA) targets Cas9 to the region immediately upstream of a 5'-NGG-3' protospacer adjacent motif (PAM) producing a double stranded break 3 bases upstream of the PAM. EnGen Spy Cas9 HF1 contains Simian virus 40 (SV40) T antigen nuclear localization sequence (NLS) on the N- and C-termini of the protein.
New England Biolabs, Inc. Nt.BsmAI – 500 units
A restriction endonuclease that recognizes the sequence GTCTCN^N.
New England Biolabs, Inc. PvuII-HF – 25000 units
A high fidelity restriction endonuclease that recognizes the sequence CAG^CTG. High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design. NEB extensively performs quality controls on all standard and high-fidelity (HF) restriction enzymes.
New England Biolabs, Inc. PstI-HF – 50000 units
A high fidelity restriction endonuclease that recognizes the sequence C_TGCA^G. High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design. NEB extensively performs quality controls on all standard and high-fidelity (HF) restriction enzymes.
New England Biolabs, Inc. T7 DNA Ligase – 750000 units
T7 DNA Ligase is an ATP-dependent ds DNA ligase from bacteriophage T7. It will catalyze the formation of a phosphodiester bond between adjacent 5' phosphate and 3' hydroxyl groups of duplex DNA. Cohesive end ligation and nick sealing can be efficiently catalyzed by T7 DNA Ligase. However, unlike T4 and T3 DNA Ligases, blunt end ligation is not efficiently catalyzed by T7 DNA Ligase. Addition of high concentrations of PEG 6000 [>= 20% (w/v)] to the reaction can force T7 DNA Ligase to have measurable activity. However, under typical reaction conditions blunt-end DNA ligation does not occur in the presence of T7 DNA Ligase, making it a good choice for applications in which blunt and cohesive ends of DNA are present but only the cohesive ends are to be joined. T7 DNA Ligase does not ligate blunt-end DNA fragments.
New England Biolabs, Inc. Exonuclease V (RecBCD) – 5000 units
Exonuclease V, a RecBCD complex from E. coli has several different enzyme activities, including an ATP-dependent single-stranded DNA endonuclease activity, ss- and ds- DNA exonuclease activity. The hydrolysis in each case is bi-directional (from both the 3' and 5' ends) and processive, producing oligonucleotides. All Exonuclease V activities have divalent cation requirements. Mg2+ is required for the exonuclease activity, while Ca2+ inhibits the exonuclease activity and allows double-stranded DNA unwinding (helicase activity) without hydrolysis.
New England Biolabs, Inc. BmtI-HF – 300 units
A high fidelity restriction endonuclease that recognizes the sequence G_CTAG^C. High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design. NEB extensively performs quality controls on all standard and high-fidelity (HF) restriction enzymes. Examples of nuclease contamination studies for some of our HF restriction enzymes are shown below.
New England Biolabs, Inc. XRN-1 – 100 units
XRN-1 is highly processive 53 exoribonuclease, requiring 5 monophosphate. It also acts on 5' monophosphate ssDNA with greatly reduced efficiency.
New England Biolabs, Inc. T4 RNA Ligase 2, truncated KQ – 2000 units
T4 RNA Ligase 2, truncated KQ (T4 Rnl2tr R55K, K227Q) specifically ligates the preadenylated 5' end of DNA or RNA to the 3' OH end of RNA. The enzyme does not use ATP for ligation but requires pre-adenylated linkers. T4 Rnl2tr R55K, K227Q is a double-point mutant of T4 RNA Ligase 2, truncated . Mutation of K227 in T4 RNA Ligase 2 reduces enzyme lysyl adenylation. K227Q reduces the formation of undesired ligation products (concatemers and circles) by T4 Rnl2tr, by reducing the trace activity of T4 Rnl2tr in transfer of adenylyl groups from linkers to the 5-phosphates of input RNAs. Mutation of R55K in T4 Rnl2tr K227Q increases the ligation activity of the enzyme to levels similar to T4 Rnl2tr. The exclusion of ATP, use of pre-adenylated linkers, and the reduced enzyme lysyl adenylation activity provide the lowest possible background in ligation reactions. This enzyme has been used for optimized linker ligation for high-throughput sequencing library construction of small RNA .