5'			Cm5	N	N	G	N9	↓		3'
3'			G	N	N	C	N13	↑		5'

SgeI can cleave DNA targets containing 5-methylcytosine on single- or double-stranded DNA

Thermo Scientific SgeI restriction enzyme recognizes m5CNNG (9/13 )^ site and cleaves optimally at 37°C in its unique buffer. For a table of enzymatic activities, double digestion conditions, and heat inactivation of this and other restriction enzymes, see Reaction conditions for restriction enzymes.

Thermo Scientific Conventional Restriction Endonucleases are a large collection of high quality restriction enzymes optimized to function in one buffer of a five-buffer system. In addition, a universal Tango buffer is provided to facilitate double enzyme digestion. All enzymes demonstrated 100% activity under recommended buffers and reaction conditions. To ensure consistent performance, Thermo Scientific Restriction Enzyme Reaction Buffers contain premixed BSA, which enhances the stability of multiple enzymes and binds contaminants that may be present in DNA preparations.

Features

• Superior quality—strict quality control and industry-leading production processes
• Convenient color-coded five-buffer system
•Includes universal Tango buffer for double digestion
•Pre-mixed BSA in reaction buffer
•Multiple restriction endonuclease specificities available

< b>Applications

•Molecular cloning
•Restriction site mapping
•Genotyping
•Southern blot
•Restriction fragment length polymorphism ( RFLP)
•SNP

Note: DNA methylated by Dcm or CpG methyltransferase will become a substrate for SgeI. More than 3-fold overdigestion with SgeI may result in incomplete digestion. For efficient cleavage, at least two copies of the SgeI recognition sequence are required. The amount of enzyme required for complete digestion of methylated DNA depends on the number of SgeI recognition sites. DNA cleavage products generated by target site cleavage can promote nonspecific cleavage of Sgel. Therefore, it is recommended to optimize the amount of enzyme used for DNA digestion. pBR322 DNA isolated from the E. coli dcm+ strain (#SD0041) was used as a DNA cleavage efficiency control. SgeI cleaves all 6 dcm methylation targets on pBR322 DNA. See product specifications for methylation sensitivity.