TatI

Product Overview

• Article Name: TatI
• Biozol Catalog Number: USB-T1027
• Supplier Catalog Number: T1027
• Alternative Catalog Number: USB-T1027-100
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-(A/T) G T A C (A/T)-3 3-(A/T) C A T G (A/T)-5 Concentration: 5u/ul Source: Thermus aquaticus CBA1-331 Form: Supplied as a liquid in 10mM Tris-HCl (pH 7.5 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. R1625 (double digests): Supplied as a liquid in 33mM Tris-acetate, (pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Incubation Temperature: Incubate at 65C. Incubate under paraffin oil in a capped vial. Incubation at 37C results in 20% activity. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 5-fold overdigestion (5u/ug lambda DNA x 1 hour) with TatI (see Star Activity). Ligation/Recutting Assay: After 2-fold overdigestion (2u/ug DNA x 1 hour) with TatI, more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 0.2uM. More than 95% of these can be recut. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours. Star Activity: An excess of enzyme (7.5u/ug DNA x 1 hour) may result in star activity. Stability during Prolonged Incubation: A minimum of 0.2 units of enzymes is required for complete digestion of 1ug of lambda DNA in 16 hours at 65C. Thermal Inactivation: Enzyme is not inactivated by incubation at 80C for 20min. Compatible Ends: Acc65I, BshNI, Bsp1407I, Pfl23II Number of Recognition Sites in DNA: Lambda: 24 PhiX174: 0 M13mp18/19: 5 pBR322: 2 pUC18/19: 2 pUC57: 2 pTZ19R/U: 1 Methylation Effects on Digestion: Dam, Dcm, CpG, EcoKI: No effect EcoBI: Effect not determined Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 65C in 50ul of assay buffer. Inactivation Procedure: To prepare prove for electrophoresis: -Stop the reaction by adding 2ul of 0.5M EDTA, pH 8.0, to achieve 20mM final concentration. Mix thoroughly, add an electrophoresis loading dye & apply to gel. To prepare DNA suitable for further enzymatic reactions: -Extract with phenol/chloroform, precipitate with ethanol or isopropanol, wash the pellet with 75% cold ethanol and air dry, -Dissolve DNA in nuclease free H2O or TE buffer, or a buffer suitable for further applications. -Check the concentration of DNA in the solution.