Thermo Scientific™ PacI

Descrizione

pJET1 DNA/SdaI fragments with inserted PacI recognition site digested with PacI, 1% agarose, 2 cleavage sites

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.

5'...T A A T▵T A A...3'

3'...A T▵T A A T T...5'

Conditions for 100% Activity:

• 1X Buffer PacI: 10mM Bis-Tris Propane-HCl (pH 6.5 at 37°C), 10mM MgCl₂ and 0.1mg/mL BSA
• Incubate at 37°C

Storage Buffer:

• PacI is supplied in: 10mM potassium phosphate (pH 7.5 at 25°C), 200mM NaCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% (v/v) glycerol

Ligation and Recleavage:

• After 50-fold overdigestion with PacI, more than 95% of the DNA fragments can be ligated and more than 95% of these can be recut

Methylation Effects:

• Dam: never overlaps —no effect
• Dcm: never overlaps —no effect
• CpG: never overlaps —no effect
• EcoKI: may overlap —effect not determined
• EcoBI: never overlaps —no effect

Digestion of Agarose-embedded DNA:

• Minimum 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded pJET1 DNA with inserted PacI recognition sequence in 16 hours

Compatible Ends:

• BstKTI, SfaNI (AsiSI), PvuI