Alo I

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.

• 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 nt away).
• The presence of SAM 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.

  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 30°C
  Double Digestion	Perform double digestion using DoubleDigest.
  Hazardous	No
  Isoschizomers	Search for commercial isoschizomers using REsearch.
  Storage Buffer	AloI 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
  		B (blue) 1X	G (green) 1X	O (orange) 1X	R (red) 1X	Tango (yellow) 1X / 2X
  Red	30°C	0-20	0-20	0-20	100	20-50	100	1X or 2X

  Methylation Effects

• Dam: may overlap - effect not determined
• Dcm: may overlap - no effect
• CpG: may overlap - cleavage impaired
• EcoKI: never overlaps - no effect
• EcoBI: never overlaps - no effect

Number of recognition sites in DNA molecules