OPTIZYME™ AarI, Fisher BioReagents™

Product Code. 11944822

Brand: Fisher BioReagents BP80691

Description

5'...C A C C T G C (N)₄ ^...3'
3'...G T G G A C G (N)₈ ^...5'

Supplied with: 10X OPTIZYME Buffer AarI, ScaI, PasI and 50X Oligonucleotide
Conditions for 100% Activity:

• 1X OPTIZYME Buffer AarI, ScaI, PasI: 10mM Bis-Tris Propane-HCl (pH 6.5 at 37°C), 10mM MgCl₂, 100mM KCl, 0.1mg/ml BSA

0.5μM of oligonucleotide

Incubate at 37°C

Enzyme Activity in OPTIZYME buffers:

• Buffer 1: No Reaction (+Oligonucleotide)

Buffer 2: No Reaction (+Oligonucleotide)

Buffer 3: 0 - 20% (+Oligonucleotide)

Buffer 4: No Reaction (+Oligonucleotide)

Buffer 5: 0 - 20% (+Oligonucleotide)

Storage Buffer:

• 10mM potassium phosphate (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% (v/v) glycerol

Ligation and Re-cleavage:

More than 95% of the DNA fragments can be ligated and re-cut after 10-fold over-digestion with AarI.

Digestion of Agarose-embedded DNA:

A minimum of 5 units of AarI is required for the digestion of 1μg of agarose-embedded lambda DNA in 16 hours.

Notes:

• At least two copies of its recognition sequence are required for cleavage with AarI
• 0.5μM oligonucleotide improves cleavage of DNAs, especially those with a single AarI site.
• Greater than 10-fold overdigestion with AarI may result in star activity.
• AarI may remain associated with cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye and SDS Solution for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis.

Methylation Effects:
Dam: Never overlaps - no effect
Dcm: Never overlaps - no effect
CpG: May overlap - cleavage impaired

• 5'...CACCTGm5C G...3'

3'...GTGGAC Gm5C...5'

(Cleavage is impaired)

EcoKI: Never overlaps - no effect

EcoBI: May overlap - effect not determined

Specifications

• Concentration: 2 U/μL
• Components: 25 to 50% Water, >50% Glycerin
• Incubator Temperature: 37°C
• pH: 7.4
• For Use With (Application): >95% of DNA fragments can be ligated and re-cut after 10-fold over-digestion with AarI
• Storage Buffer: 10mM Potassium Phosphate (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/mL BSA, 50% Glycerol
• Content And Storage: Keep container tightly closed
• Quantity: 125 U
• Cut Site: CACCTGC
• Product Type: Aarl
• Form: Liquid

Safety and Handling

SDS