Nb.Mva1269I

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

• Production of single-stranded circular DNA from supercoiled double-stranded plasmids in vitro with subsequent use in DNA sequencing, site-specific mutagenesis, etc
• Creation of nested deletions.
• Vector preparation for ligation independent cloning method
• Preparations of covalently closed, double-stranded linear DNA molecules

  Conditions for 100% Activity	1X Buffer O: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl, and 0.1 mg/mL BSA Incubate at 37°C
  Double Digestion	Perform double digestion using DoubleDigest.
  Hazardous	No
  Isoschizomers	Search for commercial isoschizomers using REsearch.
  Nicking and Cleavage	Incubation of 10 U of enzyme with 1 µg pUC19 DNA (lacking the recognition sequence of Mva1269I) for 16h at 37°C in 50 µL reaction buffer results in more than 3% conversion to circular form. Incubation of 10 U of enzyme with 1 µg pBR322 DNA for 16h at 37°C in 50 µL reaction buffer results in more tha 1% conversion to linear form.
  Storage Buffer	Nb.Mva1269I is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 50 mM NaCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/mL BSA, 50% 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
  O (Orange)	37°C	0-20	20-50	100	100	20-50	50-100	1X or 2X