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FastDigest® PmlI (Eco72I)*

* FastDigest® PmlI (Eco72I) is a proprietary formulation of Eco72I, an isoschizomer of PmlI having the same recognition and cleavage specificity. 5'-C A C^G T G-3' 3'-G T G^C A C-5'		0.7% agarose Digested lambda DNA in 5 min with FastDigest® PmlI (Eco72I) fast digest, fast digestion, time saver
#FD0364 Supplied with: 10X FastDigest® buffer	200 reactions   1 ml
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #FD0364 MSDS (English) MSDS (English-USA) MSDS (German) Chart (in pdf, 561 KB) Brochure (in pdf, 383 KB) LabAid™ (in pdf, 97 KB)

 

Features

• All FastDigest® enzymes work in the same reaction conditions.
• Single and double digestion of DNA in only 5 min.
• No star activity in prolonged incubations.
• Enhanced performance in one-hour DNA cleavage reactions.

Unit Definition
One FastDigest® Unit (FDU) is the amount of the enzyme required to cleave 1 µg of lambda DNA in 5 min at 37°C in 1X FastDigest® buffer.

Concentration
1 FDU/µl.

Recommended buffer	Reaction conditions
	Temperature	Time
FastDigest® buffer	37°C	5 min

Thermal Inactivation
FastDigest® PmlI (Eco72I) is inactivated by incubation at 80°C for 10 min.

Digestion of Plasmid DNA
1 µl of FastDigest® PmlI (Eco72I) digests up to 1 µg of plasmid DNA in 5 min, see Protocol for Fast Digestion and Analysis of Plasmid DNA with FastDigest® Enzymes and Protocol for Fast Vector Preparation with FastDigest® Enzymes.

Digestion of PCR Products
1 µl of FastDigest® PmlI (Eco72I) digests ~0.2 µg of PCR product in 5 min, see Protocol for Fast Digestion of PCR Products with FastDigest® Enzymes.

Digestion of Genomic DNA
1 µl of FastDigest® PmlI (Eco72I) digests 1 µg of genomic DNA in 5 min, or 5 µg of genomic DNA in 30 min, see Protocol for Fast Digestion of Genomic DNA with FastDigest® Enzymes.

Ligation and Recleavage
After overdigestion with 1 µl of FastDigest® PmlI (Eco72I) for 1 hour, more than 80% of the digested DNA fragments can be ligated in the reaction mixture containing 20-40 u of T4 DNA ligase/1 µg of fragments and 10% PEG. More than 90% of these can be recut.

Methylation Effects
Dam: Dcm: CpG: EcoKI: EcoBI:	never overlaps  never overlaps  completely overlaps  may overlap  may overlap		- no effect. - no effect. - blocked. - effect not determined. - effect not determined.

Recommendations for Double and Triple Digestions with FastDigest® Enzymes

Perform 5 minute double or triple digestions with FastDigest® PmlI (Eco72I) and any other FastDigest® enzyme(s) in FastDigest® buffer, see Protocol for Double Digestion with FastDigest® Enzymes and Protocol for Triple Digestion with FastDigest® Enzymes.

Updated vasario 12, 2009 13:04