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FastDigest® BsrBI (MbiI)*

* FastDigest® BsrBI (MbiI) is a proprietary formulation of MbiI, an isoschizomer of BsrBI having the same recognition and cleavage specificity. 5'-G A G^C G G-3' 3'-C T C^G C C-5'		1.4% agarose Digested lambda DNA in 5 min with FastDigest® BsrBI (MbiI) fast digest, fast digestion, time saver
#FD1274 Supplied with: 10X FastDigest® buffer	100 reactions   1 ml
Related Documents (in pdf, ~55 KB): Certificate of Analysis: #FD1274 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
FastDigest® buffer	37°C	5 min

Thermal Inactivation
FastDigest® BsrBI (MbiI) is inactivated by incubation at 65°C for 5 min.

Digestion of Plasmid DNA
1 µl of FastDigest® BsrBI (MbiI) 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® BsrBI (MbiI) 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® BsrBI (MbiI) 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® BsrBI (MbiI) for 1 hour, approximately 80% of the DNA fragments can be ligated. No more than 50% of these can be recut due to asymmetric recognition sequence of BsrBI (MbiI). The remaining uncleaved ligation products may be cut by Cfr42I (SacII) and Ecl136II (EcoICRI).

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

Recommendations for Double and Triple Digestions with FastDigest® Enzymes

Perform 5 minute double or triple digestions with FastDigest® BsrBI (MbiI) 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 29, 2008 13:27