New England Biolabs, Inc.
Established in 1975 as a private cooperative of experienced scientists, New England Biolabs is a world leader in the production of restriction endonucleases and related products for recombinant DNA technology. NEB has consistently maintained a position at the forefront of this field, and has successfully linked enzyme production efficiency to basic research in the cloning and overexpression of restriction/modification enzyme systems. This enables NEB to introduce substantial cuts in reagent costs and unsurpassed improvements in product quality and purity. Presently, NEB supplies more than 230 restriction enzymes, over 150 of which are available in recombinant form, as well as numerous recombinant polymerases and modifying enzymes for a wide variety of applications.

Why choose a recombinant enzyme?
Purity, Consistency, Affordability.

Purity
Once an enzyme system is cloned, choice of expression vector and strain background allows tight control over the production environment. For restriction endonucleases, this eliminates enzymes known to contaminate native preparations. For example, when producing AvrII from the native strain Anabaena variabilis, great care must be taken to eliminate AvrI. Similarly, when producing HaeIII from Haemophilus aegyptius, the enzyme must be purified free of HaeII. Choice of background strain also plays a key role in eliminating nonspecific endonucleases and exonucleases. Although recombinant enzymes and native enzymes are manufactured to meet the same rigorous quality control standards, it is recombinant enzymes that produce a more pure product with less processing time.

Consistency
Typically, the yields obtained for recombinant and overexpressed enzymes are significantly larger than those produced by native strains. Production of larger lots means greater product consistency and less lot-to-lot variation. Further, this large lot capability is ideal for customers that have a need for bulk quantities of enzymes. It simplifies their quality assurance procedures by reducing the time and effort normally spent to qualify enzymes supplied from different lots.

Affordability
At NEB, the introduction of recombinant enzymes has resulted in lower $/unit charges. Recombinant enzymes with lower $/unit cost allows our customers to experience substantial savings while benefiting from improved purity and consistency of product.

Expertise
NEB focuses on the analysis of restriction/modification systems at the molecular level. Our goal is to understand the regulation of these specialized systems and how they interact with DNA. This expertise is available to our customers as part of the NEB commitment to customer service.

Advances
Cloning does more than increase product quality and purity. There are many examples where native strains do not produce sufficient levels of desirable enzymes. For example, cloning of enzyme systems such as AvrII, BcgI, BspHI, HgaI, KasI, NgoMIV, has led to these being commercially available. Also, recombinant enzymes are easier to manipulate at the genetic level often leading to the commercialization of new enzymes with useful biochemical properties. For example, the thermal stable enzyme Vent_R® DNA Polymerase has a 3´ - 5´ exonuclease. Cloning of the Vent_R® DNA Polymerase gene led to the production of an exo¯ version of the enzyme that is now widely preferred for DNA sequencing.

Recombinant Enzymes From NEB:

A | B | C | D | E | F | H | I | K | M | N | P | R | S | T | X | Z |    single letter code,   icon description

Enzyme	Supplied NEBuffer	Sequence	Properties
AatII	NEBuffer 4	GACGT/C
Acc65I	NEBuffer 3	G/GTACC
AccI	NEBuffer 4	GT/MKAC
AclI	NEBuffer 4	AA/CGTT
AcuI	NEBuffer 2	CTGAAG(16/14)
AfeI	NEBuffer 4	AGC/GCT
AflII	NEBuffer 2	C/TTAAG
AflIII	NEBuffer 3	A/CRYGT
AgeI	NEBuffer 1	A/CCGGT
AhdI	NEBuffer 4	GACNNN/NNGTC
AluI	NEBuffer 2	AG/CT
AlwI	NEBuffer 4	GGATC(4/5)
ApaI @25°C	NEBuffer 4	GGGCC/C
ApaLI	NEBuffer 4	G/TGCAC
ApeKI @75°C	NEBuffer 3	G/CWGC
ApoI @50°C	NEBuffer 3	R/AATTY
AscI	NEBuffer 4	GG/CGCGCC
AseI	NEBuffer 3	AT/TAAT
AsiSI	NEBuffer 3	GCGAT/CGC
AvaI	NEBuffer 4	C/YCGRG
AvaII	NEBuffer 4	G/GWCC
AvrII	NEBuffer 2	C/CTAGG
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BamHI	NEBuffer 3	G/GATCC
BanI	NEBuffer 4	G/GYRCC
BbvCI	NEBuffer 4	CCTCAGC(-5/-2)
BbvI	NEBuffer 2	GCAGC(8/12)
BccI	NEBuffer 1	CCATC(4/5)
BcgI	NEBuffer 3	(10/12)CGANNNNNNTGC(12/10)
BciVI	NEBuffer 4	GTATCC(6/5)
BclI @50°C	NEBuffer 3	T/GATCA
BglI	NEBuffer 3	GCCNNNN/NGGC
BglII	NEBuffer 3	A/GATCT
BlpI	NEBuffer 4	GC/TNAGC
BmrI	NEBuffer 2	ACTGGG(5/4)
BmtI	NEBuffer 2	GCTAG/C
Bpu10I	NEBuffer 3	CCTNAGC(-5/-2)
BsaAI	NEBuffer 3	YAC/GTR
BsaI @50°C	NEBuffer 3	GGTCTC(1/5)
BseRI	NEBuffer 2	GAGGAG(10/8)
BseYI	NEBuffer 3	CCCAGC(-5/-1)
BsiWI @55°C	NEBuffer 3	C/GTACG
BslI @55°C	NEBuffer 3	CCNNNNN/NNGG
BsmAI @55°C	NEBuffer 3	GTCTC(1/5)
BsmBI @55°C	NEBuffer 3	CGTCTC(1/5)
BsmFI @65°C	NEBuffer 4	GGGAC(10/14)
BsmI @65°C	NEBuffer 2	GAATGC(1/-1)
BsoBI	NEBuffer 2	C/YCGRG
BspEI	NEBuffer 3	T/CCGGA
BspHI	NEBuffer 4	T/CATGA
BspMI	NEBuffer 3	ACCTGC(4/8)
BspQI @50°C	NEBuffer 3	GCTCTTC(1/4)
BsrFI	NEBuffer BsrFI	R/CCGGY
BssHII @50°C	NEBuffer 3	G/CGCGC
BssSI	NEBuffer 3	CACGAG(-5/-1)
BstEII @60°C	NEBuffer 3	G/GTNACC
BstXI	NEBuffer 3	CCANNNNN/NTGG
BstXI	NEBuffer 3	CCANNNNN/NTGG
BstYI @60°C	NEBuffer 2	R/GATCY
BtsCI @50°C	NEBuffer 4	GGATG(2/0)
BtsI @55°C	NEBuffer 4	GCAGTG(2/0)
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ClaI	NEBuffer 4	AT/CGAT
CspCI	NEBuffer 2	(11/13)CAANNNNNGTGG(12/10)
CviAII @25°C	NEBuffer 4	C/ATG
CviKI-1	NEBuffer 4	RG/CY
CviQI @25°C	NEBuffer 3	G/TAC
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DdeI	NEBuffer 3	C/TNAG
DpnI	NEBuffer 4	GA/TC
DpnII	NEBuffer DpnII	/GATC
DraI	NEBuffer 4	TTT/AAA
DraIII	NEBuffer 3	CACNNN/GTG
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EaeI	NEBuffer 1	Y/GGCCR
EagI	NEBuffer 3	C/GGCCG
EarI	NEBuffer 1	CTCTTC(1/4)
EcoO109I	NEBuffer 4	RG/GNCCY
EcoP15I	NEBuffer 3	CAGCAG(25/27)
EcoRI	NEBuffer EcoRI	G/AATTC
EcoRV	NEBuffer 3	GAT/ATC
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FatI @55°C	NEBuffer 2	/CATG
FauI @55°C	NEBuffer 1	CCCGC(4/6)
Fnu4HI	NEBuffer 4	GC/NGC
FokI	NEBuffer 4	GGATG(9/13)
FseI	NEBuffer 4	GGCCGG/CC
FspI	NEBuffer 4	TGC/GCA
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HaeII	NEBuffer 4	RGCGC/Y
HaeIII	NEBuffer 2	GG/CC
HgaI	NEBuffer 1	GACGC(5/10)
HhaI	NEBuffer 4	GCG/C
HincII	NEBuffer 3	GTY/RAC
HindIII	NEBuffer 2	A/AGCTT
HinfI	NEBuffer 2	G/ANTC
HinP1I	NEBuffer 2	G/CGC
HpaI	NEBuffer 4	GTT/AAC
HpaII	NEBuffer 1	C/CGG
HphI	NEBuffer 4	GGTGA(8/7)
Hpy166II	NEBuffer 4	GTN/NAC
Hpy188I	NEBuffer 4	TCN/GA
Hpy188III	NEBuffer 4	TC/NNGA
Hpy99I	NEBuffer 4	CGWCG/
HpyAV	NEBuffer 4	CCTTC(6/5)
HpyCH4III	NEBuffer 4	ACN/GT
HpyCH4IV	NEBuffer 1	A/CGT
HpyCH4V	NEBuffer 4	TG/CA
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I-CeuI	NEBuffer 4	CGTAACTATAACGGTCCTAAGGTAGCGAA(-9/-13)
I-SceI	NEBuffer I-SceI	TAGGGATAACAGGGTAAT(-9/-13)
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KasI	NEBuffer 2	G/GCGCC
KpnI	NEBuffer 1	GGTAC/C
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MboI	NEBuffer 3	/GATC
MboII	NEBuffer 2	GAAGA(8/7)
McrBC	NEBuffer 2
MfeI	NEBuffer 4	C/AATTG
MluI	NEBuffer 3	A/CGCGT
MlyI	NEBuffer 4	GAGTC(5/5)
MmeI	NEBuffer 4	TCCRAC(20/18)
MnlI	NEBuffer 2	CCTC(7/6)
MscI	NEBuffer 4	TGG/CCA
MseI	NEBuffer 2	T/TAA
MslI	NEBuffer 2	CAYNN/NNRTG
MspA1I	NEBuffer 4	CMG/CKG
MspI	NEBuffer 2	C/CGG
MwoI @60°C	NEBuffer 3	GCNNNNN/NNGC
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NaeI	NEBuffer 1	GCC/GGC
Nb.BbvCI	NEBuffer 2	CCTCAGC
Nb.BsmI @65°C	NEBuffer 3	GAATGC
Nb.BsrDI @65°C	NEBuffer 2	GCAATG
Nb.BtsI	NEBuffer 4	GCAGTG
NciI	NEBuffer 4	CC/SGG
NcoI	NEBuffer 3	C/CATGG
NdeI	NEBuffer 4	CA/TATG
NgoMIV	NEBuffer 4	G/CCGGC
NheI	NEBuffer 2	G/CTAGC
NheI-HF™	NEBuffer 4	G/CTAGC
NlaIII	NEBuffer 4	CATG/
NlaIV	NEBuffer 4	GGN/NCC
NmeAIII	NEBuffer 4	GCCGAG(21/19)
NotI	NEBuffer 3	GC/GGCCGC
NruI	NEBuffer 3	TCG/CGA
NsiI	NEBuffer 3	ATGCA/T
NspI	NEBuffer 2	RCATG/Y
Nt.AlwI	NEBuffer 2	GGATC(4/-5)
Nt.BbvCI	NEBuffer 4	CCTCAGC(-5/-7)
Nt.BspQI @50°C	NEBuffer 3	GCTCTTC(1/-7)
Nt.BstNBI @55°C	NEBuffer 3	GAGTC(4/-5)
Nt.CviPII	NEBuffer 4	(0/-1)CCD
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PacI	NEBuffer 1	TTAAT/TAA
PaeR7I	NEBuffer 4	C/TCGAG
PciI	NEBuffer 3	A/CATGT
PflMI	NEBuffer 3	CCANNNN/NTGG
PhoI @75°C	NEBuffer 3	GG/CC
PI-PspI @65°C	NEBuffer PI-PspI	TGGCAAACAGCTATTATGGGTATTATGGGT(-13/-17)
PI-SceI	NEBuffer PI-SceI	ATCTATGTCGGGTGCGGAGAAAGAGGTAAT(-15/-19)
PleI	NEBuffer 4	GAGTC(4/5)
PmeI	NEBuffer 4	GTTT/AAAC
PpuMI	NEBuffer 4	RG/GWCCY
PshAI	NEBuffer 4	GACNN/NNGTC
PsiI	NEBuffer 4	TTA/TAA
PspGI @75°C	NEBuffer 2	/CCWGG
PspOMI	NEBuffer 4	G/GGCCC
PspXI	NEBuffer 2	VC/TCGAGB
PstI	NEBuffer 3	CTGCA/G
PvuII	NEBuffer 2	CAG/CTG
PvuII-HF™	NEBuffer 4	CAG/CTG
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RsaI	NEBuffer 1	GT/AC
RsrII	NEBuffer 4	CG/GWCCG
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SacI	NEBuffer 1	GAGCT/C
SacII	NEBuffer 4	CCGC/GG
SalI	NEBuffer 3	G/TCGAC
SalI-HF™	NEBuffer 4	G/TCGAC
SapI	NEBuffer 4	GCTCTTC(1/4)
Sau96I	NEBuffer 4	G/GNCC
SbfI	NEBuffer 4	CCTGCA/GG
ScaI	NEBuffer 3	AGT/ACT
ScaI-HF™	NEBuffer 4	AGT/ACT
SfaNI	NEBuffer 3	GCATC(5/9)
SfcI	NEBuffer 4	C/TRYAG
SfiI @50°C	NEBuffer 2	GGCCNNNN/NGGCC
SfoI	NEBuffer 2	GGC/GCC
SgrAI	NEBuffer 4	CR/CCGGYG
SmaI @25°C	NEBuffer 4	CCC/GGG
SnaBI	NEBuffer 4	TAC/GTA
SpeI	NEBuffer 2	A/CTAGT
SphI	NEBuffer 2	GCATG/C
SphI-HF™	NEBuffer 4	GCATG/C
SspI	NEBuffer SspI	AAT/ATT
StuI	NEBuffer 2	AGG/CCT
StyD4I	NEBuffer 2	/CCNGG
StyI	NEBuffer 3	C/CWWGG
SwaI @25°C	NEBuffer 3	ATTT/AAAT
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TaqαI @65°C	NEBuffer 3	T/CGA
TfiI @65°C	NEBuffer 3	G/AWTC
TliI @75°C	NEBuffer 3	C/TCGAG
Tsp509I @65°C	NEBuffer 1	/AATT
TspRI @65°C	NEBuffer 4	NNCASTGNN/
Tth111I @65°C	NEBuffer 4	GACN/NNGTC
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XbaI	NEBuffer 2	T/CTAGA
XcmI	NEBuffer 2	CCANNNNN/NNNNTGG
XhoI	NEBuffer 2	C/TCGAG
XmaI	NEBuffer 4	C/CCGGG
XmnI	NEBuffer 2	GAANN/NNTTC
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ZraI	NEBuffer 1	GAC/GTC