OPTIZYME™ AloI, Fisher BioReagents™

Name: OPTIZYME&trade; AloI, Fisher BioReagents&trade;
Brand: Fisher Bioreagents
SKU: 11994822
Price: 180.00 EUR

Marca: Fisher Bioreagents BP8075-1

Dettagli aggiuntivi : Peso : 0.01000kg

Codice prodotto. 11994822

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Descrizione

5'...^ ₇(N) G A A C (N)₆ T C C (N)_12-13^...3'
3'...^_12-13(N) C T T G (N)₆ A G G (N)₇ ^...5'

Supplied with: 10X OPTIZYME Buffer 5
Conditions for 100% Activity:

1X OPTIZYME Buffer 5: 10mM Tris-HCl (pH 8.5 at 37°C), 10mM MgCl₂, 100mM KCl and 0.1mg/mL BSA
Incubate at 30°C

Enzyme Activity in OPTIZYME buffers:

Buffer 1: 0 - 20%
Buffer 2: 0 - 20%
Buffer 3: 0 - 20%
Buffer 4: 20 - 50%
Buffer 5: 100%

Storage Buffer:

10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol

Ligation and Re-cleavage:

Approximately 70% of the DNA fragments can be ligated and more than 80% of these can be re-cut after 10-fold over-digestion with AloI.

Notes:

Incubation at 37°C results in 20% activity.
AloI produces double-strand cuts on both sides from the interrupted recognition site. Its unique feature is a degenerate cleavage point on the 3 side of the recognition sequence (12 or 13 nucleotides away).
The presence of S-adenosylmethionine in the reaction mixture results in incomplete cleavage with AloI.
Greater than 10-fold overdigestion with AloI may result in star activity.
AloI may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye & SDS Solution for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis.

Methylation Effects:
Dam: May overlap - effect not determined

5'...GAAC(N)₄Gm6A TCC...3'

3'...CTTG(N)₄C Tm6AGG...5'

(Cleavage effect not determined)

Dcm: May overlap - no effect

5'...GAAC(N)₆TCm5CW GG...3'

3'...CTTG(N)₆AG GWm5CC...5'

(No effect on cleavage)

CpG: May overlap - cleavage impaired

5'...m5C GAAC(N)₆TCC...3'

3'... Gm5CTTG(N)₆AGG...5'

(Cleavage is blocked)

5'...GAAC(N)₆TCm5C G...3'

3'...CTTG(N)₆AG Gm5C...5'

(No effect on cleavage)

5'...GAAm5C G(N)₅TCC...3'

3'...CTT Gm5C(N)₅AGG...5'

(No effect on cleavage)

EcoKI: Never overlaps - no effect

EcoBI: Never overlaps - no effect

Specifica

Concentrazione	1 to 3 U/μL
Temperatura incubatore	30°C
Da utilizzare con (applicazione)	>95% of DNA fragments can be ligated and re-cut after 50-fold over-digestion with AloI
Content And Storage	Keep container tightly closed
Sito di taglio	(7/12)GAAC(N)6TCC(12/7)
Forma	Liquid

Componenti	25 to 50% Water, >50% Glycerin
pH	7.4
Tampone di conservazione	10mM Tris HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA, 50% Glycerol
Quantità	100 U
Product Type	AloI

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OPTIZYME™ AloI, Fisher BioReagents™

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