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FastDigest® DpnI

CH3 5'-G A^T C-3' 3'-C T^A G-5' CH3		1.4% agarose Digested pBR322 DNA in 5 min with FastDigest® DpnI fast digest, fast digestion, time saver
#FD1703 Supplied with: 10X FastDigest® buffer	50 reactions   1 ml
#FD1704 Supplied with: 10X FastDigest® buffer	100 reactions   1 ml
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #FD1703, #FD1704 MSDS (English) MSDS (English-USA) MSDS (German) Chart (in pdf, 834 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 pBR322 DNA (dam methylated) in 5 min at 37°C in 1X FastDigest® buffer.

Concentration
1 FDU/µl.

Recommended buffer	Reaction conditions
	Temperature	Time
1X FastDigest® buffer	37°C	5 min

Thermal Inactivation
FastDigest® DpnI is inactivated by incubation at 80°C for 5 min.

Digestion of Plasmid DNA
1 µl of FastDigest® DpnI 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
Not applicable.

Digestion of Genomic DNA
1 µl of FastDigest® DpnI digests 1 µg of genomic DNA in 10 min, or 5 µg of genomic DNA in 60 min, see Protocol for Fast Digestion of Genomic DNA with FastDigest® Enzymes.

Ligation and Recleavage
After overdigestion with 1 µl of FastDigest® DpnI for 1 hour, more than 70% of the digested pBR322 DNA fragments can be ligated and more than 95% of these can be recut.

Methylation Effects
Dam: Dcm: CpG: EcoKI: EcoBI:	does not cut dam- DNA.  never overlaps  may overlap  never overlaps  may overlap	- no effect. - no effect. - no effect. - effect not determined.

Recommendations for Double and Triple Digestions with FastDigest® Enzymes

Perform 5 minute double or triple digestions with FastDigest® DpnI 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 kovo 17, 2008 14:45