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FastDigest® Fnu4HI (SatI)*

* FastDigest® Fnu4HI (SatI) is a proprietary formulation of SatI, an isoschizomer of Fnu4HI having the same recognition and cleavage specificity. 5'-G C^N G C-3' 3'-C G N^C G-5'		1.4% agarose Digested lambda DNA in 5 min with FastDigest® Fnu4HI (SatI) fast digest, fast digestion, time saver
#FD1644 Supplied with: 10X FastDigest® buffer	20 reactions   1 ml
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #FD1644 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 lambda DNA 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® Fnu4HI (SatI) is inactivated by incubation at 65°C for 5 min.

Digestion of Plasmid DNA
1 µl of FastDigest® Fnu4HI (SatI) 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® Fnu4HI (SatI) digests ~0.2 µg of PCR product in 10 min, see Protocol for Fast Digestion of PCR Products with FastDigest® Enzymes.

Digestion of Genomic DNA
1 µl of FastDigest® Fnu4HI (SatI) 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® Fnu4HI (SatI) for 1 hour, more than more than 60% of the digested DNA fragments can be ligated in a reaction mixture containing 10-30 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  may overlap  never overlaps  never overlaps	- no effect. - no effect. - blocked. - no effect. - no effect.

Compatible Ends
GC^(C/G)GC - BcnI, Bme1390I, NciI, ScrFI.
GC^(A/T)GC - Bme1390I, MvaI, ScrFI.

Recommendations for Double and Triple Digestions with FastDigest® Enzymes

Perform 5 minute double or triple digestions with FastDigest® Fnu4HI (SatI) 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 05, 2008 15:51