HindIII

Kód produktu: ER0502 Kód výrobce: ER0502 Kód dodavatele: {5ED9D19F-54A4-4E7E-B538-8AF1DB6E6257} Výrobce: Life Technologies Czech Republic s.r.o.

`5'...A↓A G C T T...3' 3'...T T C G A↑A...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.

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 37°C
Digestion of Agarose-embedded DNA	Minimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double Digestion	Perform double digestion using DoubleDigest.
Hazardous	No
Isoschizomers	Search for commercial isoschizomers using REsearch.
Storage Buffer	HindIII is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 250 mM KCl, 1 mM DTT, 0.1 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)	37°C	0-20	20-50	0-20	100	50-100	50-100	1X or 2X

Methylation Effects

Dam: never overlaps - no effect.
Dcm: never overlaps - no effect.
CpG: never overlaps - no effect.
EcoKI: never overlaps - no effect.
EcoBI: may overlap - cleavage impaired.

Methylation type	Sequence	Cleavage effect
EcoBI (`TGA(N)₈TGCT`)	5'...`TGm6AAGCTT(N)₃ TGCT`...3' 3'...`AC TTCGAA(N)₃m6ACGA`...5'	Impaired

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
6	0	1	1	1	1
pTZ19R/U	pTZ57R	pBluescriptIIKS(-/+)	pBluescriptIISK(-/+)	pACYC177	pACYC184
1	1	1	1	1	1

New sites generated by ligation

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
`A^AGCTT`	`AAGCTAGCTT`	[AluI/FastDigest AluI] BspOI (BmtI)/FastDigest BmtI (BspOI) Cac8I [CviJI] FspBI (BfaI)/FastDigest BfaI (FspBI) NheI/FastDigest NheI [SetI]

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5'...A↓A G C T T...3' 3'...T T C G A↑A...5

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