Thermo Scientific™ Van91I (PflMI) (10 U/μL)

Descrizione

Lambda DNA digested with Van91I (PflMI), 0.7% agarose, 14 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'..C C A NN N N^N T G G...3'

3'..G G T N^N N NN A C C...5'

Conditions for 100% Activity

• 1X Buffer R: 10mM Tris-HCl (pH 8.5 at 37°C), 10mM MgCl₂, 100mM KCl and 0.1mg/ml BSA
• Incubate at 37°C

Storage Buffer

• Van91I is supplied in: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 Recleavage

• After 50-fold overdigestion with Van91I, more than 90% of DNA fragments can be ligated and recut

Methylation Effects

• Dam: never overlaps – no effect
• Dcm: may overlap – blocked
• CpG: never overlaps – no effect
• EcoKI: never overlaps – no effect
• EcoBI: never overlaps – no effect&

Digestion of Agarose-embedded DNA

• Minimum 5 units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours

Note:

Van91I is blocked by overlappingdcm methylation. To avoiddcm methylation, use adam,dcm- strain such as GM2163 (#M0099).