Recombinant Enzymes from NEB
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1 New England Biolabs, Inc. Tel: (odes) Tel: (suppot) Fax: Recombinant Enzymes fom NEB New England Biolabs, Inc. Established in 1975 as a pivate coopeative of expeienced scientists, New England Biolabs is a wold leade in the poduction of estiction endonucleases and elated poducts fo ecombinant DNA technology. NEB has consistently maintained a position at the foefont of this field, and has successfully linked enzyme poduction efficiency to basic eseach in the cloning and oveexpession of estiction/modification enzyme systems. This enables NEB to intoduce substantial cuts in eagent costs and unsupassed impovements in poduct quality and puity. Pesently, NEB supplies moe than 230 estiction enzymes, ove 150 of which ae available in ecombinant fom, as well as numeous ecombinant polymeases and modifying enzymes fo a wide vaiety of applications. Why choose a ecombinant enzyme? Puity, Consistency, Affodability. Puity Once an enzyme system is cloned, choice of expession vecto and stain backgound allows tight contol ove the poduction envionment. Fo estiction endonucleases, this eliminates enzymes known to contaminate native pepaations. Fo example, when poducing Av II fom the native stain Anabaena vaiabilis, geat cae must be taken to eliminate Av I. Similaly, when poducing Hae III fom Haemophilus aegyptius, the enzyme must be puified fee of Hae II. Choice of backgound stain also plays a key ole in eliminating nonspecific endonucleases and exonucleases. Although ecombinant enzymes and native enzymes ae manufactued to meet the same igoous quality contol standads, it is ecombinant enzymes that poduce a moe pue poduct with less pocessing time. Consistency Typically, the yields obtained fo ecombinant and oveexpessed enzymes ae significantly lage than those poduced by native stains. Poduction of lage lots means geate poduct consistency and less lot-to-lot vaiation. Futhe, this lage lot capability is ideal fo customes that have a need fo bulk quantities of enzymes. It simplifies thei quality assuance pocedues by educing the time and effot nomally spent to qualify enzymes supplied fom diffeent lots. Affodability At NEB, the intoduction of ecombinant enzymes has esulted in lowe $/unit chages. Recombinant enzymes with lowe $/unit cost allows ou customes to expeience substantial savings while benefiting fom impoved puity and consistency of poduct. Expetise NEB focuses on the analysis of estiction/modification systems at the molecula level. Ou goal is to undestand the egulation of these specialized systems and how they inteact with DNA. This expetise is available to ou customes as pat of the NEB commitment to custome sevice. Advances Cloning does moe than incease poduct quality and puity. Thee ae many examples whee native stains do not poduce sufficient levels of desiable enzymes. Fo example, cloning of enzyme systems such as Av II, Bcg I, BspH I, Hga I, Kas I, NgoM IV, has led to these being commecially available. Also, ecombinant enzymes ae easie to manipulate at the genetic level often leading to the commecialization of new enzymes with useful biochemical popeties. Fo example, the themal stable enzyme Vent DNA Polymease has a 3 5 exonuclease. Cloning of the Vent DNA Polymease gene led to the poduction of an exo vesion of the enzyme that is now widely pefeed fo DNA sequencing. (see othe side fo a complete listing of ecombinant enzymes available fom New England Biolabs) (see othe side) Technical Bulletin Cloning (4/24/06)
2 Recombinant Enzymes fom NEB (continued) Restiction Endonucleases Aat II #R0117S #R0117L 2, AsiS I #R0630S #R0630L 2, BsaA I #R0531S #R0531L 2, Acc I #R0161S #R0161L 2, Ava I #R0152S #R0152L BseR I #R0581S #R0581L Acl I #R0598S #R0598L 300 units 1, Ava II #R0153S #R0153L BseY I #R0635S #R0635L 2, Acu I #R0641S #R0641L Av II #R0174S #R0174L BsiW I #R0553S #R0553L 300 units 1, Afl II #R0520S #R0520L BamH I #R0136S #R0136L Bsl I #R0555S #R0555L Afl III #R0541S #R0541L 1, Ban I #R0118S #R0118L 2 Bsm I #R0134S #R0134L 2, Age I #R0552S #R0552L 1, Bbv I #R0173S #R0173L 150 units 750 units BsmB I #R0580S #R0580L Ahd I #R0584S #R0584L BbvC I #R0601S #R0601L BsoB I #R0586S #R0586L Alu I #R0137S #R0137L Bcc I #R0704S #R0704L BspE I #R0540S #R0540L Alw I #R0513S #R0513L 1, Bcg I #R0545S #R0545L 1, BspH I #R0517S #R0517L 2, Apa I #R0114S #R0114L Bcl I #R0160S #R0160L 1 BspM I #R0502S #R0502L ApaL I #R0507S #R0507L 2, 12, Bgl I #R0143S #R0143L BsF I #R0562S #R0562L 2, ApeK I #R0643S #R0643L 1, Bgl II #R0144S #R0144L BssH II #R0199S #R0199L Apo I #R0566S #R0566L Asc I #R0558S #R0558L 2, 2, Blp I #R0585S #R0585L Bm I #R0600S #R0600L 2, 25 units 125 units BssS I #R0587S #R0587L BstE II #R0162S #R0162L Ase I #R0526S #R0526L Bsa I #R0535S #R0535L BstY I #R0523S #R0523L Page 2 (Cloning)
3 New England Biolabs, Inc. Tel: (odes) Tel: (suppot) Fax: Recombinant Enzymes fom NEB (continued) BtsC I #R0647S #R0647L EcoR I #R0101S #R0101L Hpa II #R0171S #R0171L Cla I #R0197S #R0197L EcoR V #R0195S #R0195L 4,000 units 20,000 units Hph I #R0158S #R0158L 400 units CspC I #R0645S #R0645L 2, Fnu4H I #R0178S #R0178L Hpy99 I #R0615S #R0615L CviA II #R0640S #R0640L Fok I #R0109S #R0109L Hpy188 I #R0617S #R0617L CviK I-1 #R0710S #R0710L 1, Fse I #R0588S #R0588L Hpy188 III #R0622S #R0622L 2, Dde I #R0175S #R0175L 2, Fsp I #R0135S #R0135L 2, HpyCH4 III #R0618S #R0618L 1, Dpn I #R0176S #R0176L Hae II #R0107S #R0107L HpyCH4 IV #R0619S #R0619L 2, Dpn II #R0543S #R0543L Hae III #R0108S #R0108L 1 HpyCH4 V #R0620S #R0620L Da I #R0129S #R0129L Hga I #R0154S #R0154L Kas I #R0544S #R0544L 1, Da III #R0510S #R0510L Hha I #R0139S #R0139L Kpn I #R0142S #R0142L 1 Eae I #R0508S #R0508L Hinc II #R0103S #R0103L Mbo I #R0147S #R0147L 2, Eag I #R0505S #R0505L 2, Hind III #R0104S #R0104L Mbo II #R0148S #R0148L 1, Ea I #R0528S #R0528L 2, Hinf I #R0155S #R0155L 2 Mfe I #R0589S #R0589L 2, EcoO109 I #R0503S #R0503L HinP1 I #R0124S #R0124L Mlu I #R0198S #R0198L EcoP15 I #R0646S #R0646L 25 units 125 units Hpa I #R0105S #R0105L 2, Mly I #R0610S #R0610L (see othe side) Page 3 (Cloning)
4 Recombinant Enzymes fom NEB (continued) Mme I #R0637S #R0637L Nsp I #R0602S #R0602L 1, Sap I #R0569S #R0569L 50 units Mnl I #R0163S #R0163L 2, Pac I #R0547S #R0547L 1, Sau96 I #R0165S #R0165L Msc I #R0534S #R0534L 1, PaeR7 I #R0177S #R0177L Sbf I #R0642S #R0642L 1, Mse I #R0525S #R0525L 2, PflM I #R0509S #R0509L Sca I #R0122S #R0122L Msp I #R0106S #R0106L 2 Pho I #R0705S #R0705L Sfi I #R0123S #R0123L MspA1 I #R0577S #R0577L 2, Ple I #R0515S #R0515L 1, Sfo I #R0606S #R0606L Mwo I #R0573S #R0573L 1, Pme I #R0560S #R0560L 2, SgA I #R0603S #R0603L Nae I #R0190S #R0190L 2, PpuM I #R0506S #R0506L 2, Sma I #R0141S #R0141L Nci I #R0196S #R0196L PshA I #R0593S #R0593L SnaB I #R0130S #R0130L 2, Nco I #R0193S #R0193L PspG I #R0611S #R0611L Spe I #R0133S #R0133L 2, Nde I #R0111S #R0111L 4,000 units 20,000 units Pst I #R0140S #R0140L Sph I #R0182S #R0182L 1, NgoM IV #R0564S #R0564L Pvu II #R0151S #R0151L 2 Ssp I #R0132S #R0132L Nhe I #R0131S #R0131L Rsa I #R0167S #R0167L Sty I #R0500S #R0500L 1 Nla III #R0125S #R0125L 2, Rs II #R0501S #R0501L StyD4 I #R0638S #R0638L Nla IV #R0126S #R0126L Sac I #R0156S #R0156L Swa I #R0604S #R0604L Not I #R0189S #R0189L 2, Sac II #R0157S #R0157L Taq I #R0149S #R0149L 4,000 units 20,000 units Nsi I #R0127S #R0127L Sal I #R0138S #R0138L Tfi I #R0546S #R0546L 2, Page 4 (Cloning)
5 New England Biolabs, Inc. Tel: (odes) Tel: (suppot) Fax: Recombinant Enzymes fom NEB (continued) Tli I #R0612S #R0612L Tsp509 I #R0576S #R0576L TspR I #R0582S #R0582L Tth111 I #R0185S 400 units #R0185L Xba I #R0145S #R0145L 1 Xcm I #R0533S #R0533L 2, Xho I #R0146S #R0146L 2 Xma I #R0180S #R0180L 1, Xmn I #R0194S #R0194L N.Alw I #R0627S #R0627L 2, N.BstNB I #R0607S #R0607L N.BbvC IA #R0631S #R0631L N.BbvC IB #R0632S #R0632L McBC #M0272S #M0272L 2, Homing Endonucleases I-Ceu I #R0699S #R0699L 1, I-Sce I #R0694S #R0694L 1, PI-Psp I #R0695S #R0695L PI-Sce I #R0696S #R0696L DNA Methylases Alu I Methylase #M0220S #M0220L BamH I Methylase #M0223S #M0223L dam Methylase #M0222S #M0222L 2, EcoR I Methylase #M0211S #M0211L Hae III Methylase #M0224S #M0224L 2, Hha I Methylase #M0217S #M0217L Hpa II Methylase #M0214S #M0214L Human DNA (cytosine-5) Methyltansfease (Dnmt1) #M0230S 50 units #M0230L Msp I Methylase #M0215S #M0215L CpG Methylase (M.Sss I) #M0226S #M0226L Taq I Methylase #M0219S #M0219L DNA Polymeases phi29 DNA Polymease #M0269S #M0269L 1, Taq DNA Polymease with ThemoPol Buffe #M0267S 400 units #M0267L Taq DNA Polymease with Standad Taq Buffe #M0273S 400 units #M0273L Taq 2X Maste Mix #M0270S 100 eactions #M0270L 500 eactions Quick-Load Taq 2X Maste Mix #M0271S 100 eactions #M0271L 500 eactions Taq PCR Kit #E5000S 200 eactions Taq PCR Kit with Contols #E5100S 200 eactions 9 N m DNA Polymease #M0260S #M0260L Theminato DNA Polymease #M0261S #M0261L Bst DNA Polymease, Lage Fagment #M0275S 1,600 units #M0275L 8,000 units Vent DNA Polymease #M0254S #M0254L Vent (exo ) DNA Polymease #M0257S #M0257L Deep Vent DNA Polymease #M0258S #M0258L Deep Vent (exo ) DNA Polymease #M0259S #M0259L T7 DNA Polymease #M0256S 300 units #M0256L 1, (see othe side) Page 5 (Cloning)
6 Recombinant Enzymes fom NEB (continued) M-MuLV Revese Tansciptase #M0253S #M0253L DNA Polymease I (E. coli) #M0209S #M0209L 2, DNA Polymease I, L. (Klenow) Fag. #M0210S #M0210L Klenow Fagment (3 5 exo ) #M0212S #M0212L SP6 RNA Polymease #M0207S #M0207L T4 DNA Polymease #M0203S 150 units #M0203L 750 units T7 RNA Polymease #M0251S #M0251L 2 Teminal Tansfease #M0252S #M0252L 2, Phusion High-Fidelity DNA Polymease #F-530S #F-530L Phusion Hot Stat High-Fidelity DNA Polymease #F-540S #F-540L DyNAzyme EXT DNA Polymease #F-505S #F-505L DyNAzyme II Hot Stat DNA Polymease #F-504S #F-504L DNA Ligases T4 DNA Ligase #M0202S 20,000 units #M0202L 100,000 units Themus aquaticus (Taq) DNA Ligase #M0208S #M0208L T4 RNA Ligase #M0204S #M0204L Nucleases Exonuclease I (E. coli) #M0293S #M0293L 1 Exonuclease III (E. coli) #M0206S #M0206L 2 Lambda Exonuclease #M0262S #M0262L T7 Exonuclease #M0263S #M0263L RecJ f #M0264S #M0264L 1, Exonuclease T #M0265S #M0265L 1, RNases ShotCut RNase III #M0245S 130 units #M0245L 650 units RNase H #M0297S #M0297L 1, RNase I f #M0243S #M0243L RNase Inhibito M0307S M0307L DNA Repai Poteins T7 Endonuclease I #M0292S #M0292L 1, Endonuclease III (Nth) #M0268S #M0268L Endonuclease IV #M0304S #M0304L 1, Endonuclease V #M0305S #M0305L 1, Endonuclease VIII #M0299S #M0299L 2, Topoisomease I (E. coli) #M0301S #M0301L Uacil-DNA Glycosylase (UDG) #M0280S 400 units #M0280L USER Enzyme #M5505S #M5505L Fpg #M0240S #M0240L hogg1 #M0241S #M0241L APE 1 #M0282S #M0282L 50 units 400 units 80 units 400 units Single-standed DNA Binding Poteins RecA #M0249S 200 µg #M0249L 1,000 µg T4 Gene 32 Potein #M0300S 100 µg #M0300L 500 µg Recombinase/Tansposase Ce Recombinase #M0298S 50 units #M0298L TnsABC* Tansposase #P7190S 10 µl (10 xns) Antactic Phosphatase Antactic Phosphatase #M0289S #M0289L 2, Agaase β-agaase I #M0392S #M0392L DNA Kinase T4 Polynucleotide Kinase #M0201S #M0201L 2, Page 6 (Cloning)
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