Thermo Scientific™ PdiI (NaeI)
The PdiI (NaeI) restriction enzyme recognizes GCC^GGC sites and cuts best at 37°C in Tango buffer (Isoschizomers: MroNI, NaeI, NgoMIV).
Manufacturer: Thermo Scientific™ ER1522
pBR322 DNA digested with PdiI (NaeI), 0.7% agarose, 4 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'...G C C▵G G C...3'
3'...C G G▵C C G...5'
Conditions for 100% Activity:
- 1X Buffer Tango: 33mM Tris-acetate (pH 7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/mL BSA
- Incubate at 37°C
- PdiI is supplied in: 10mM Tris-HCl (pH 7.5 at 25°C), 500mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mL BSA, 0.15% Triton X-100 and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with PdiI, more than 90% of the DNA fragments can be ligated and recut
- Dam: never overlaps —no effect
- Dcm: never overlaps —no effect
- CpG: completely overlaps —blocked
- EcoKI: never overlaps —no effect
- EcoBI: never overlaps —no effect
Digestion of Agarose-embedded DNA:
- Minimum 20units of the enzyme are required for complete digestion of 1μg of agarose-embedded pBR322 DNA in 16hours
Certain sites in pBR322 are difficult to cleave with PdiI, the same as with its prototype NaeI
Assayed using pBR322 DNA (#SD0041)