Restriction endonucleses Turbo

Description

In response to customer demand, we have launched a new line of Turbo Restriction Endonucleases designed to save time during DNA digestion workflows.

Our team has evaluated selected enzymes for complete digestion in 15 minutes, optimizing enzyme concentration and reaction conditions. The current Turbo Restriction Endonuclease lineup includes 76 enzymes.

The unified Turbo protocol uses 1 μl of enzyme to digest 1 μg of DNA. For best results, highly purified DNA preparations should be used, especially PCR fragments. Please note that complete digestion of some PCR products, as well as cleavage of supercoiled plasmid DNA by Turbo enzymes, may require longer incubation times than those specified in the protocol.

Important! Turbo formats are supplied primarily with ROSE Buffer (see activity table).

Turbo enzymes should not be diluted and must be used in their original, undiluted form.

List of enzymes

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Protocols

Turbo enzymes have been tested for their ability to digest DNA in a short period of time. During testing, 1 μl of a Turbo enzyme was added to a reaction mixture containing 1 μg of DNA (phage or plasmid) and 1X universal ROSE Buffer (for some enzymes, their own optimal buffer was used). After incubation for 15 minutes, complete DNA digestion was observed.

Standard Fast Digestion Protocol:

For a 20 μl reaction mixture:
10X SE Buffer ROSE (or optimal buffer) – 2 μl
DNA substrate – 0.2–1 μg
BSA (10 mg/ml) (optional)* – 0.2 μl
Sterile water – to 20 μl
+ 1 μl Turbo Restriction Endonuclease (or 1 μl each of two Turbo enzymes for double digestion).

Carefully mix the reaction. Incubate at the optimal temperature for 10 or 15 minutes.

*Add to the reaction if BSA is required for maximal enzyme activity.

Note:

Turbo enzymes must be used only in their original, undiluted form.

Double Digestion of DNA with Turbo Restriction Endonucleases

This protocol is intended for double digestion of DNA using Turbo Restriction Endonucleases.

If simultaneous digestion with two restriction enzymes is required, we recommend using universal ROSE Buffer (B021), or any restriction enzyme buffer in which both enzymes exhibit at least 50% activity. Otherwise, sequential digestion with each enzyme separately is recommended, with inactivation of the first enzyme prior to the second digestion. Information on restriction enzyme activity in ROSE Buffer and other reaction buffers can be found in the Service / Restriction Enzyme Activity section.

Reaction Optimal SibEnzyme (ROSE) Buffer is a specially developed universal reaction buffer for restriction endonucleases. The vast majority of restriction enzymes exhibit 50–100% activity in this buffer.

Standard Turbo Double Digestion Protocol:

For a 20 μl reaction mixture:

Sterile water – to 20 μl
Optimal or universal reaction buffer (10X) – 2 μl;
DNA substrate – 0.2 to 1 μg;
BSA (10 mg/ml) (optional)* – 0.2 μl;
+ 1 μl Restriction Enzyme #1 and 1 μl Restriction Enzyme #2

Incubate at the optimal temperature for 0.5–1 hour.

*BSA should be added if at least one of the enzymes requires it for maximal activity.

Note:

The final glycerol concentration in the digestion reaction should not exceed 5% to avoid star activity. Therefore, no more than a total of 2 μl of enzymes should be added to a standard 20 μl reaction mixture.
If the enzymes have different optimal reaction temperatures, sequential digestion is recommended (starting with the lower-temperature enzyme), with inactivation of the first enzyme before the second digestion.

Activity table

Restrictions endonucleases Turbo

№	Enzyme	Prototype	Recognition site	Cat No	Buffer	Reaction temp, ^oC (activity in % at..)
№	Enzyme	Prototype	Recognition site	Cat No	Buffer
1	AatII	AatII	GACGT^C	E287T/ E288T	ROSE	37
2	Acc16I	MstI	TGC^GCA	E001T/ E002T	ROSE	37
3	Acc65I	Acc65I, KpnI^	G^GTACC	E003T/ E004T	ROSE	37
4	AccB7I	PflMI	CCANNNN^NTGG	E179T/ E180T	ROSE	37
5	AccBSI	BsrBI	GAG^CGG	E007T/E008T	ROSE	37
6	AcsI	ApoI	R^AATTY	E013T/ E014T	ROSE	50
7	AfeI	Eco47III	AGC^GCT	E213T/ E214T	ROSE	37
8	AhlI	SpeI	A^CTAGT	E173T/ E174T	ROSE	37
9	AluI	AluI	AG^CT	E015T/ E016T	ROSE	37
10	ApaI	ApaI	GGGCC^C	E019T/ E020T	ROSE	37
11	AsiGI	AgeI	A^CCGGT	E235T/ E236T	ROSE	37
12	AspA2I	AvrII	C^CTAGG	E245T/ E246T	W+	37
13	AspLEI	HhaI	GCG^C	E221T/ E222T	ROSE	37
14	AspS9I	Sau96I	G^GNCC	E117T/ E118T	ROSE	37
15	AsuNHI	NheI	G^CTAGC	E063T/ E064T	Y+	37
16	BamHI	BamHI	G^GATCC	E021T/ E022T	ROSE	37
17	BglI	BglI	GCCNNNN^NGGC	E025T/ E026T	ROSE	37
18	BglII	BglII	A^GATCT	E027T/ E028T	ROSE	37
19	Bme18I	AvaII	G^GWCC	E029T/ E030T	ROSE	37
20	BmtI	NheI^	GCTAG^C	E457T/ E458T	ROSE	37
21	Bpu14I	AsuII	TT^CGAA	E033T/ E034T	ROSE	37
22	Bsa29I	ClaI	AT^CGAT	E205T/ E206T	ROSE	37
23	Bso31I	Eco31I, BsaI	GGTCTC(1/5)	E285T/ E286T	ROSE+	55
24	Bsp19I	NcoI	C^CATGG	E047T/ E048T	2W+	37
25	BstACI	AcyI	GR^CGYC	E093T/ E094T	ROSE	37
26	BstAUI	Bsp1407I	T^GTACA	E267T/ E268T	ROSE	37
27	BstDEI	BstDEI	C^TNAG	E227T/ E228T	ROSE	60
28	BstNSI	NspI	RCATG^Y	E251T/ E252T	ROSE	37
29	BstV2I	BbvII	GAAGAC(2/6)	E297T/ E298T	ROSE	55
30	CciI	BspHI	T^CATGA	E565T/ E566T	ROSE	55
31	CciNI	NotI	GC^GGCCGC	E203T/ E204T	ROSE	37
32	DraI	AhaIII	TTT^AAA	E055T/ E056T	ROSE	37
33	DraIII	DraIII	CACNNN^GTG	E309T/ E310T	ROSE	37
34	DriI	Eam1105I	GACNNN^NNGTC	E193T/ E194T	ROSE	37
35	EcoRI	EcoRI	G^AATTC	E057T/ E058T	ROSE	37
36	EcoRV	EcoRV	GAT^ATC	E059T/ E060T	ROSE	37
37	FauNDI	NdeI	CA^TATG	E009T/ E010T	ROSE	37
38	Fsp4HI	Fnu4HI	GC^NGC	E095T/ E096T	ROSE	37
39	HaeIII	HaeIII	GG^CC	E067T/ E068T	ROSE	37
40	HindII	HindII	GTY^RAC	E201T/ E202T	ROSE	37
41	HindIII	HindIII	A^AGCTT	E073T/ E074T	ROSE	37
42	HinfI	HinfI	G^ANTC	E075T/ E076T	ROSE	37
43	HpaI	HpaI	GTT^AAC	E077T/ E078T	Y	37
44	HpaII	HpaII	C^CGG	E161T/ E162T	ROSE	37
45	HspAI	HhaI^	G^CGC	E069T/ E070T	ROSE	37
46	KpnI	KpnI	GGTAC^C	E079T/ E080T	ROSE+	37
47	MfeI	MfeI	C^AATTG	E295T/ E296T	ROSE+	37
48	MluI	MluI	A^CGCGT	E085T/ E086T	ROSE	37
49	MnlI	MnlI	CCTC(7/6)	E481T/ E482T	ROSE+	37
50	MspI	HpaII	C^CGG	E091T/ E092T	ROSE	37
51	PceI	StuI	AGG^CCT	E105T/ E106T	ROSE	50 (at 37 – 50%)
52	PciI	BspLU11I	A^CATGT	E275T/ E276T	ROSE	37
53	PsiI	PsiI	TTA^TAA	E279T/ E280T	ROSE	37
54	Psp124BI	SacI	GAGCT^C	E107T/ E108T	G	37
55	PspCI	PmaCI	CAC^GTG	E475T/ E476T	B+	37
56	PspN4I	NlaIV	GGN^NCC	E089T/ E090T	ROSE	37
57	PspXI	PspXI	VC^TCGAGB	E477T/ E478T	ROSE	37
58	PstI	PstI	CTGCA^G	E109T/ E110T	ROSE	37
59	PvuII	PvuII	CAG^CTG	E111T/ E112T	ROSE+	37
60	RsaI	RsaI	GT^AC	E113T/ E114T	ROSE	37
61	SalI	SalI	G^TCGAC	E115T/ E116T	O	37
62	SfaNI	SfaNI	GCATC(5/9)	E165T/ E166T	O	37
63	SfiI	SfiI	GGCCNNNN^NGGCC	E123T/ E124T	ROSE+	50 (at 37 – 75%)
64	Sfr274I	XhoI	C^TCGAG	E125T/ E126T	ROSE	50 (at 37 – 75%)
65	Sfr303I	SacII	CCGC^GG	E127T/ E128T	ROSE	37
66	SmaI	SmaI	CCC^GGG	E177T/ E178T	Y	37
67	SmiI	SwaI	ATTT^AAAT	E225T/ E226T	O+	37
68	SphI	SphI	GCATG^C	E129T/ E130T	ROSE	37
69	Sse9I	Tsp509I	^AATT	E217T/ E218T	ROSE	55 (at 37 – 75%)
70	SspI	SspI	AAT^ATT	E041T/ E042T	ROSE+	37
71	TaqI	TaqI	T^CGA	E133T/ E134T	ROSE	65
72	Tru9I	MseI	T^TAA	E199T/ E200T	ROSE	65
73	VneI	ApaLI	G^TGCAC	E137T/ E138T	ROSE	37
74	VspI	VspI	AT^TAAT	E139T/ E140T	ROSE	37
75	XbaI	XbaI	T^CTAGA	E141T/ E142T	ROSE	37
76	ZrmI	ScaI	AGT^ACT	E005T/ E006T	ROSE	37