Modifying Enzymes
New England Biolabs, Inc. Thermostable 5' App DNA/RNA Ligase – 10 reactions
Thermostable 5' App DNA/RNA Ligase is a point mutant of catalytic lysine of RNA ligase from Methanobacterium thermoautotrophicum. This enzyme is ATP independent. It requires a 5' pre-adenylated linker for ligation to the 3' -OH end of either RNA or single stranded DNA (ssDNA). The enzyme is also active in ligation of RNA with 2' -O-methylated 3 end to 5-adenylated linkers. The optimal temperature for ligation reaction is 60-65°C. The mutant ligase is unable to adenylate the 5' -phosphate of RNA or ssDNA, which reduces the formation of undesired ligation products (concatemers and circles). The ability of the ligase to function at 65°C might reduce the constraints of RNA secondary structure in RNA ligation experiments.
New England Biolabs, Inc. T4 RNA Ligase 2, truncated – 10000 units
T4 RNA Ligase 2' truncated (T4 Rnl2'truncated) specifically ligates the pre-adenylated 5' end of DNA or RNA to the 3 end of RNA. The enzyme does not require ATP for ligation but does need the pre-adenylated substrate. T4 Rnl2'truncated is expressed from a plasmid in E. coli which encodes the first 2'9 amino acids of the full length T4 RNA Ligase 2' Unlike the full length ligase, T4 Rnl2'truncated is unable to adenylate the 5' end of the substrate, and as a result it cannot ligate the phosphorylated 5' end of RNA or DNA to the 3' end of RNA . This enzyme, also known as Rnl2'(1-2'9) has been used for optimized linker ligation for the cloning of microRNAs. This enzyme reduces background ligation because it can only use adenylated primers).
New England Biolabs, Inc. Uracil-DNA Glycosylase (UDG) – 1000 units
E. coli Uracil-DNA Glycosylase (UDG) catalyses the release of free uracil from uracil-containing DNA. UDG efficiently hydrolyzes uracil from single-stranded or double-stranded DNA, but not from oligomers (6 or fewer bases).
New England Biolabs, Inc. α1-6 Mannosidase - 800 units
a1-6 Mannosidase is a highly specific exoglycosidase that removes unbranched a1-6 linked mannose residues from oligosaccharides. When used in conjunction with a1-2,3 Mannosidase, the a1-6 Mannosidase will cleave a1-6 mannose residues from branched carbohydrate substrates.
New England Biolabs, Inc. ZraI – 200 units
A restriction endonuclease that recognizes the sequence GAC_^GTC.
New England Biolabs, Inc. Mung Bean Nuclease – 7500 units
Mung Bean Nuclease is a single-strand specific DNA and RNA endonuclease which will degrade single-stranded extensions leaving blunt, ligatable ends.
New England Biolabs, Inc. AluI Methyltransferase – 100 units
AluI Methyltransferase modifies the cytosine residue (C5) of the sequence AGCT.
New England Biolabs, Inc. CpG Methyltransferase (M.SssI) – 100 units
The CpG Methyltransferase, M.SssI, methylates all cytosine residues (C5) within the double-stranded dinucleotide recognition sequence 5'...CG...3'
New England Biolabs, Inc. SalI-HF – 2000 units
A high fidelity restriction endonuclease that recognizes the sequence G^TCGAC. 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. MscI – 1250 units
A restriction endonuclease that recognizes the sequence TGG_^CCA.
New England Biolabs, Inc. PNGase F, Recombinant – 75000 units
PNGase F is the most effective enzymatic method for removing almost all N-linked oligosaccharides from glycoproteins. PNGase F, Recombinant is a recombinant amidase, which cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides.
New England Biolabs, Inc. Bacteroides Heparinase II – 80 units
Bacteroides Heparinase II (also called Heparin Lyase II) is cloned from Bacteroides eggerthii. It is a low specificity enzyme that is active on both heparin and heparan sulfate. Bacteroides Heparinase II cleaves the glycosidic bond between N-sulfated and glucuronic or iduronic acid residues. The reaction yields oligosaccharide products containing unsaturated uronic acids which can be detected by UV spectroscopy at 232 nm. When used alone this enzyme rarely yields complete depolymerization of a polysaccharide chain, however disaccharide analysis is enhanced when used in combination with Heparinase I and III.
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 .
