Brand: Across International
Publisher: ACROSS INTERNATIONAL LLC
Details: Storage: −20°C UNSPSC Code: 12352200 Components: Enzyme Solution; SuRE/Cut Buffer H 10x concentrated RIDADR: NONH for all modes of transport Analysis Note: Activity in PCR buffer: 0% General description: Not I recognizes the sequence GCGG*C*CG°C and generates fragments with 5′-cohesive termini. Not I belongs to the class of "rare-cutter" enzymes. It is one of the two known enzymes that recognize an octameric sequence comprised solely of G and C residues. Contents: •Not I •SuRE/Cut Buffer H (10x) Other Notes: For life science research only. Not for use in diagnostic procedures. Quality: Absence of nonspecific endonuclease activities 1 μg Ad2 DNA is incubated for 16 hours in 50 μl SuRE/Cut Buffer H with an excess of Not I. The number of enzyme units which do not change the enzyme-specific pattern is stated in the certificate of analysis. Absence of exonuclease activity Approximately 5 μg [ 3 H] labeled calf thymus DNA are incubated with 3 μl Not I for 4 hours at +37°C in a total volume of 100 μl 50 mM Tris-HCl, 10 mM MgCl 2 , 1 mM Dithioerythritol, pH approximately 7.5. Under these conditions, no release of radioactivity is detectable, as stated in the certificate of analysis. Specifications: Average size of fragment generated Prokaryotic genomic DNA: Not I fragments are between 20 and 1,000 kb, depending on the GC content. Yeast genomic DNA: Not I fragments are, on average, 200 kb. Mammalian genomic DNA: Not I fragments are approximately 1,000 kb. Compatible ends Not I ends are compatible with ends generated by Eae I and EclX I (Xma III). Isoschizomers The enzyme has no known isoschizomers. Methylation sensitivity Not I is inhibited by the presence of 5-
