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MultiTaq (5 U/µl)
Technical data
Source: Purified from an E. Coli strain carrying Thermus species DNA Polymerase overproducing plasmid.
Analysis condition: 25 mM Tris•HCl pH 9.0 at 25°C, 50mM KCl, 2 mM MgCl2, 0.1mg/ml gelatin, 200 µM each dATP, dGTP, dTTP, 100 µM [α32-P]dCTP (0.05µCi/nmol), 12.5 µg activated salmon sperm DNA.
Unit definition: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmol of dNTP into an acid-insoluble form in 30 minutes at 74ºC.
Storage & Dilution buffer: 50% glycerol (v/v), 20 mM Tris•HCl pH 8.7 at 25ºC, 100 mM KCl, 0.1 mM EDTA and stabilizers.
Associated activities: The enzyme has a 5’ to 3’ polymerisation-dependent exonuclease replacement activity but lacks a 3’ to 5’ exonuclease activity. The enzyme has "extendase activity" allowing TA cloning.
Application & Quality control: Primer extension. The enzyme is free of nicking and priming activities, exonucleases and unspecific endonucleases. SDS/PAGE - 95 kD band, >98% pure. Activity and stability tested via thermocycling. The error rate per nucleotide per cycle is ~8.3x10-5; the accuracy is ~ 1.2x104. Estimated half life at 95ºC is 1.5 hours.
Reaction Buffers
• 10x Reaction buffer D (Mg2+ free): 800mM Tris•HCl pH 9.4-9.5 at 25ºC, 200 mM (NH4)2SO4, 0.2% w/v Tween-20.
• 10x Reaction buffer DF (Mg2+ and detergent free): pH 9.4-9.5 800mM Tris•HCl, 200 mM (NH4)2SO4.
Routine storage of Reaction Buffers and 25 mM MgCl2 at -20ºC is recommended.
DO NOT store Buffers at 4ºC!
Operating conditions: It is recommended to add the components in the following order: 10xbuffer, H2O, MgCl2 final concentration 2.0-2.5 mM (2.5mM is strongly recommended), dNTPs (200uM), DNA template (5ng-100ng/100µl), primers 30-100pmol/100µl) and finally the polymerase (2-5 U/100µl), for hard templates up to 10U/100ul.
Shipping & Storage conditions:
Shipping and storage at room temperature for up to 1 month has no detrimental effects on the quality of MultiTaq.
Routine storage: -20ºC
Safety warnings and precautions: This product and its components should be handled only by persons trained in laboratory techniques. It is advisable that suitable protective clothing, such as laboratory overalls, safety glasses and gloves be worn. Care should be taken to avoid contact with skin or eyes. In case of contact wash immediately with water.
Some applications of this product may be protected and require a license which is not provided by the purchase of this product. US DNA recommends all users to obtain licenses as necessary.
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