Tru1I (MseI)

Kód produktu: ER0981 Kód výrobce: ER0981 Kód dodavatele: {35038F49-22D2-4529-A4E4-0A974065280E} Výrobce: Life Technologies Czech Republic s.r.o.

`5'...T↓T A A...3' 3'...A A T↑T...5'`

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Detailní popis

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Highlights

Superior quality – stringent quality control and industry leading manufacturing process
Convenient color-coded Five Buffer System
Includes universal Tango buffer for double-digestions
BSA premixed in reaction buffers
Wide selection of restriction endonuclease specificities

Applications

Molecular cloning
Restriction site mapping
Genotyping
Southern Blot
Restriction fragment length polymorphism (RFLP)
SNP

Note

For methylation sensitivity refer to product specifications.

Compatible Ends	Csp6I, FspBI, NdeI, VspI.
Conditions for 100% Activity	1X Buffer R: 10 mM Tris-HCl (pH 8.5 at 37°C), 10 mM MgCl₂, 100 mM KCl, and 0.1 mg/mL BSA Incubate at 65°C To ensure higher efficiency of digestion, perform the cleavage reaction under paraffin oil
Double Digestion	Perform double digestion using DoubleDigest.
Hazardous	No
Isoschizomers	Search for commercial isoschizomers using REsearch.
Note	Incubation at 37°C results in 10% activity. We recommend using Tru1I, HC at 37°C.
Storage Buffer	Tru1I is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM KCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/mL BSA, and 50% (v/v) glycerol.
Storage Condition	-20 C

Reaction conditions

Recommended buffer for 100% activity	Optimal temperature	Enzyme activity in Thermo Scientific buffers, %						Tango buffer for double digestion
Recommended buffer for 100% activity	Optimal temperature	B (blue) 1X	G (green) 1X	O (orange) 1X	R (red) 1X	Tango (yellow) 1X / 2X		Tango buffer for double digestion
R (Red)	65°C	50-100	50-100	20-50	100	50-100	100	1X or 2X

Methylation Effects

Dam: never overlaps – no effect.
Dcm: never overlaps – no effect.
CpG: never overlaps – no effect.
EcoKI: may overlap – blocked.
EcoBI: never overlaps – no effect.

Methylation type	Sequence	Cleavage effect
EcoKI`(AAC(N)₆GTGC)`	5'...`TTAm6AC(N)₆G TGC`...3' 3'...`AAT TG(N)₆Cm6ACG`...5'	Blocked

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1	2	3	4	5
0	0-20		50-100

Number of recognition sites in DNA molecules

Lambda	ΦX174	M13mp18/19	pBR322	puc18/19	pUC57
195	35	63	15	13	13
pTZ19R/U	pTZ57R	pBluescriptIIKS(-/+)	pBluescriptIISK(-/+)	pACYC177	pACYC184
18	18	19	19	16	12

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition Sequence	Second Enzyme	Enzymes recognizing newly generated recognition sequence
`T^TAA`	NdeI/FastDigest NdeI (`CA^TATG`)	FaiI
`T^TAA`	FastDigest MseI (SaqAI) (`T^TAA`) VspI (AseI)/FastDigest AseI (VspI) (`AT^TAAT`)	FastDigest MseI (SaqAI) Tru1I (MseI)/FastDigest Tru1I

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition Sequence	Newly generated sequence after reaction	Enzymes that cleave the newly generated sequence
`T^TAA`	`TTATAA`	AanI (PsiI)/FastDigest PsiI (AanI) FaiI

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5'...T↓T A A...3' 3'...A A T↑T...5'

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