对应每一种限制性内切酶，New England Biolabs 均提供彩色盖子的10X NEBuffer，以保证100%活性。下面表格中列出每一种酶及其随酶提供的最适宜的NEBuffer。同时也列出每种酶在四种不同的NEBuffer中的大致活性，以帮助选择双酶切时最适宜的环境。除非特别指出，温育温度均为37°C。

注意事项：表格中所列出的酶活性是近似值，计算酶活性时所采用的底物与计算该酶的单位时所采用的底物相同。 缓冲液在使用前应充分解冻，并混合均匀。(http://www.ebiotrade.com/)

NEB限制酶在各种NEBuffer 中的活性(A-C)

Enzyme	Supplied NEBuffer	% Activity in NEBuffers
		1	2	3	4
Dde I	3	75	100	100	75
Dpn I	4	100	100	75	100
Dpn II	U	NR	NR	dd100	NR
Dra I	4	75	75	50	100
Dra III	3 + BSA	100	75	100	25
Drd I	4	25	50	10	100
Eae I	1	100	100	50	100
Eag I	3	10	25	100	10
Ear I	1	100	100	50	100
Eci I	2 + BSA	50	100	100	50
EcoN I	4	100	100	75	100
EcoO109 I	4 + BSA	100	100	75	100
EcoR I	Udd	100	100	100	100
EcoR V	3 + BSA	50	75	100	50
Fat I *	SE-G	10	100	50	50
Fau I * @55°C	SE-B	100	25	10	50
Fnu4H I	4	10	25	25	100
Fok I	4	50	100	75	100
Fse I	4 + BSA	100	75	0	100
Fsp I	4	10	75	10	100
Hae II	4 + BSA	75	100	50	100
Hae III	2	50	100	25	75
Hga I	1	100	75	50	100
Hha I	4 + BSA	75	100	100	100
Hinc II	3 + BSA	75	100	100	100
Hind III	2	50	100	10	50
Hinf I	2	75	100	75	75
HinP1 I	2	100	100	100	100
Hpa I	4	25	50	10	100
Hpa II	1	100	50	10	100
Hph I	4	NR	75	0	100
Hpy99 I	4 + BSA	10	10	0	100
Hpy188 I	4	50	25	10	100
Hpy188 III	4 + BSA	100	100	10	100
HpyCH4III	4	100	50	25	100
HpyCH4IV	1	100	25	10	25
HpyCH4V	4	50	50	25	100
I-Ceu I	4 + BSA	10	10	0	100
I-Sce I	U + BSA	10	50	50	50
Kas I	2 + BSA	75	100	75	75
Kpn I	1 + BSA	100	75	0	50
Mbo I	3	75	100	100	100
Mbo II	2	100	100	50	100
McrBC	2 + BSA + GTP	50	100	10	75
Mfe I	4	75	50	10	100
Mlu I	3	25	75	100	50
Mly I	4 + BSA	50	50	25	100
Mme I	4 + SAM	NR	NR	NR	100
Mnl I	2 + BSA	75	100	50	75
Msc I	4	75	75	75	100
Mse I	2 + BSA	75	100	75	75
Msl I	2	100	100	25	100
Msp I	2	75	100	50	50
MspA1 I	4 + BSA	50	100	50	100
Mwo I @60°C	U	10	75	100	75
Nae I	1	100	75	10	75
N.Alw I	2	25	100	25	50
Nar I	1	100	75	75	75
N.BbvC IA	2	50	100	10	100
N.BbvC IB	2	50	100	10	100
N.BstNB I @55°C	U	0	10	10	10
Nci I	4	100	25	10	100
Nco I	4	100	100	100	100
Nde I	4	75	100	75	100
NgoM IV	4	100	50	10	100
Nhe I	2 + BSA	100	100	10	100
Nla III	4 + BSA	25	25	25	100
Nla IV	4 + BSA	10	10	10	100
Not I	3 + BSA	0	75	100	50
Nru I	U	0	10	75	10
Nsi I	U	25	25	50	10
Nsp I	2 + BSA	100	100	0	100
Pac I	1 + BSA	100	75	10	100
PaeR7 I	4	25	100	10	100
Pci I *	SE-O	25	75	100	50
PflF I	4 + BSA	0	25	25	100
PflM I	3 + BSA	0	100	100	50
PI-Psp I @65°C	U + BSA	0	10	10	10
PI-Sce I	U + BSA	0	0	0	0
Ple I	4	100	100	75	100
Pme I	4 + BSA	0	50	10	100
Pml I	1 + BSA	100	75	0	75
PpuM I	4	0	25	0	100
PshA I	4 + BSA	50	75	10	100
Psi I *	SE-B	50	100	0	25
PspG I @75°C	2	75	100	50	100
PspOM I *	SE-Y	25	100	50	100
Pst I	3 + BSA	75	75	100	50
Pvu I	3 + BSA	10	75	100	10
Pvu II	2	100	100	100	100
Rsa I	1	100	100	50	100
Rsr II	4	25	75	10	100
Sac I	1 + BSA	100	50	10	100
Sac II	4	25	75	10	100
Sal I	U + BSA	0	0	50	0
Sap I	4	75	50	0	100
Sau3A I	U + BSA	100	50	10	100
Sau96 I	4	50	100	100	100
Sbf I *	SE-Y	75	50	0	100
Sca I	U	25	75	dd50	25
ScrF I	4	100	100	100	100
SexA I	3 + BSA	25	25	100	100
SfaN I	3	0	75	100	25
Sfc I	4 + BSA	75	50	10	100
Sfi I @50°C	2 + BSA	0	100	10	100
Sfo I	2	25	100	50	50
SgrA I	4	100	50	10	100
Sma I @25°C	4	0	0	0	100
Sml I @55°C	4 + BSA	25	75	25	100
SnaB I	4 + BSA	25	50	25	dd100
Spe I	2 + BSA	75	100	25	75
Sph I	2	100	100	50	100
Ssp I	U	50	100	50	50
Stu I	2	100	100	50	100
Sty I	3 + BSA	25	75	100	10
StyD4 I	2	10	100	100	100
Swa I @25°C	3 + BSA	10	10	100	10
Taq I @65°C	U + BSA	50	75	75	75
Tfi I @65°C	3	100	100	dd100	100
Tli I @75°C	3 + BSA	25	25	100	10
Tse I @65°C	3	75	100	100	75
Tsp45 I @65°C	1 + BSA	100	25	0	75
Tsp509 I @65°C	1	100	100	100	NR
TspR I @65°C	4 + BSA	25	50	25	100
Tth111 I @65°C	1	100	25	25	100
Xba I	2 + BSA	0	100	75	75
Xcm I	2	10	100	50	50
Xho I	2 + BSA	75	100	100	100
Xma I	4 + BSA	25	50	0	100
Xmn I	2 + BSA	100	100	75	100
Zra I *	SE-B	100	100	25	100