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限制酶Restriction Enzyme (Jena Bio)

Restriction enzyme	Tris-HCl(Tris-Acetate) {Bis Tris Propane-HCl}		NaCl(K-Acetate) {KCl}	MgCl₂ (Mg-Acetate)	DTT	BSA	TX-100	Temp.	Buffer	包裝
Unit	mM	pH (25℃)	mM	mM	mM	mg/ml	%	℃
Acc I	(20)	7.9	(50)	(10)	1	100	─	37	B5	300U
Alu I	10	7.9	─	10	1	100	─	37	B1	600U
Asu II	10	7.9	50	10	1	100	0.1	37	B2*	3500U
BamH I	10	7.9	100	5	1	100	─	37	U	10000U
Bcl I	10	7.9	50	10	1	100	─	50	B2	2500U
Bgl I	100	7.9	50	5	─	100	0.025	37	U	2000U
Bgl II	50	7.9	100	10	1	100	─	37	B3	1300U
BseA I	10	8	100	5	1	100	0.02	55	U	650U
BseB I	10	7.9	50	10	1	100	─	60	B2	4500U
BsC I	50	7.9	100	10	1	100	─	55	B3	800U
BshF I	(20)	7.9	(50)	(10)	1	100	─	37	B5	7000U
BsiS I	(33)	7.9	(66)	(10)	0.5	100	0.1	55	U	2200U
BssA I	20	8.5	{100}	3	─	100	0.04	65	U	300U
BstE II	10	7.4	{100}	5	1	100	0.1	60	U	2000U
CspA I	{10}	7.0	─	10	1	100	─	37	U	150U
Dpn I	(33)	7.9	(66)	(10)	─	100	─	37	U	250U
EcoR I	100	7.4	50	5	─	100	0.025	37	U	5000U
EcoR V	10	7.9	50	10	1	100	─	37	B2	3000U
Hind III	10	7.9	50	10	1	100	─	37	B2	8000U
Hinf I	50	7.9	100	10	1	100	─	37	B3	3300U
Hpa I	(20)	7.9	(50)	(10)	1	100	─	37	B5	1000U
Kpn I	10	7.0	─	10	1	100	0.01	37	U	3500U
Mbo I	10	8.0	{100}	10	1	100	─	37	U	300U
MspC I	10	7.9	150	10	1	100	─	37	B4	1300U
Nae I	10	7.9	─	10	1	100	─	37	B1	350U
Nco I	50	7.9	100	10	1	100	0.02	37	B3*	600U
Nhe I	(20)	7.9	(50)	(10)	1	100	─	37	B5	550U
Not I	50	7.9	100	5	1	100		37	BU	300U
Nru I	50	8.0	{100}	10	─	100	─	37	U	700U
PspP I	10	7.9	50	10	1	100	─	25	B2	900U
Pst I	50	7.4	100	10	1	100	─	37	U	8000U
Pvu II	10	7.9	50	10	1	100	─	37	B2	4500U
Rsa I	10	7.9	50	10	1	100	─	37	B2	1000U
Sal I	10	7.9	150	10	1	100	─	37	B4	2000U
Sau3A I	10	7.9	50	10	1	100	─	37	B2	500U
Sca I	10	7.4	100	10	1	100	─	37	U	1200U
Sfi I	10	7.9	50	10	1	100	─	50	B2	550U
SgrB I	10	7.9	─	10	1	100	0.1	37	B1*	1600U
Sla I	10	7.9	150	10	1	100	─	37	B4	5000U
Sma I	(20)	7.9	(50)	(10)	1	100	─	25	B5	1100U
SnaB I	{10}	7.0	─	10	1	100	─	37	U	350U
Sph I	10	7.9	50	10	1	100	─	37	B2	250U
SseB I	50	7.9	100	10	1	100	─	37	B3	2000U
Ssp I	50	7.9	100	10	1	100	─	37	B3	600U
Sst I	10	7.9	─	10	1	100	─	37	B1	1600U
Sty I	50	7.9	100	10	1	100	─	37	B3	6000U
Taq I	20	8.5	{100}	3	─	100	0.04	65	U	4500U
Xba I	10	7.9	50	10	1	100	─	37	B2	3500U

Restriction Enzymes Buffer Guide

According to assay conditions the restriction endonucleases were divided into five groups. Each group is more active in one of the five following reaction buffers:

The activity of each restriction endonuclease was evaluated in each of the above five buffers containing 100 ug/ml bovine serum albumin (BSA).

Notes: BSA is not included into the supplied reaction buffers and should be added separately from the supplied stock solution (1 mg/ml). Some restriction endonucleases require Triton X-100 (TX-100). This means that 100 % of the activity is obtained using this additive. Several enzymes - BamH I, Bgl I, BseA I, BsiS I, BssA I, BstE II, CspA I, EcoR I, Kpn I, Mbo I, Nru I, Pst I, Sca I, SnaB I, and Taq I require unique (U) buffers for optimal reaction conditions. The composition of each unique buffer is presented in specific restriction endonuclease descriptions, also in the Technical Data Sheet provided with each enzyme.

10x reaction buffers should be thawed completely and mixed thoroughly before using.

Double Digestion of DNA with Jena Bioscience Enzymes

Simultaneous cleavage of DNA with two different restriction endonucleases is a common time-saving procedure. The best buffer can be chosen following the Enzyme Activity Chart which rates the activity of each restriction endonuclease in the 5 Jena Bioscience buffers.

In the table below you will find recommended buffers for double digestions using 15 of the most common restriction endonucleases. If no single Jena Bioscience buffer can be found to satisfy the buffer requirements of both enzymes, the reactions can be done sequentially (seq). First, cleave with the restriction endonuclease that requires the lower salt reaction conditions, then adjust the salt concentration of the reaction to approximate the reaction conditions of the second restriction endonuclease. Add the second enzyme and incubate to complete the second reaction.

When using restriction endonucleases in non-optimal buffers, more enzyme or longer digestion time may be needed to compensate for the slower rate of cleavage under those conditions.

All the reactions were carried out in the presence of BSA (100 ug/ml). Our experience indicates that it is important to use BSA in reaction mixtures in order to obtain successful digestions of DNA. The presence of BSA gives complete and reproducible cleavages for a range of DNA substrates. BSA stabilizes the enzymes when digestions are perfomed for more than one hour at 37°C, since many restriction endonucleases in reaction buffers without BSA can survive at this temperature for 10-20 minutes only or even less. Also, BSA binds metal ions, and other chemicals, which might be present in buffers or DNA preparations, thereby inactivating restriction endonucleases.

Note: BSA is not included into the supplied reaction buffers and should be added separately from the supplied stock solution (1 mg/ml). * Requires Triton X-100 for optimal activity. TX-100 is included into the supplied reaction buffer. The following enzymes can exhibit "star" activity: BamH I, Bcl I, BseB I, BssA I, EcoR I, EcoR V, Hind III, Hpa I, Kpn I, Nco I, Nru I, Pst I, Pvu II, Sal I, Sca I, SnaB I, Sph I, Ssp I, Xba I.

台灣轉錄科技有限公司 分生器材網

台灣轉錄科技有限公司分生器材網