Taq Polymerase. Data sheet. Order No.: BS (For research and in vitro applications only) Batch No.: Best before: Bio&SELL

Transcription

1 Data sheet Order No.: BS Order No.: BS units 500 units (For research and in vitro applications only) Batch No.: Best before: Appearance: clear liquid Colour: transparent 1

2 Description Taq Polymerase - robust, versatile and economical. The Taq-DNA-Polymerase achieves excellent results in your PCR applications because you can optimize each of your PCR reactions individually. Included in the delivery of Taq Polymerase are two different buffer systems (with and without detergent), separate MgCl 2 solution and a solution for GC-rich templates and for minimizing background. Select the optimal buffer system for the particular application. With the detergent-containing Buffer B, the yields of the PCR reaction can be optimized. The BD Buffer without Tween-20 is recommended for detergentsensitive, downstream applications. Scope of delivery: Taq Polymerase (5 U/µl) (! red cap) Reaction Buffer B (10x) (! blue cap) Reaction Buffer BD (10x) (! green cap) MgCl 2 Solution (25 mm) (! grey cap) Solution S (10x) (! yellow cap) 2

3 Reaction Buffers and Solution S 10x Reaction Buffer B (free of Mg 2+ ): 800 mm Tris HCl (ph 9,4-9,5 at 25ºC), 200 mm (NH 4 ) 2 SO 4, 0,2% w/v Tween-20 10x Reaction Buffer BD (free of Mg 2+ and detergents): 800 mm Tris HCl (ph 9,4-9,5 at 25ºC), 200 mm (NH 4 ) 2 SO 4 Note: Routine storage of Reaction Buffers B and BD and of 25 mm MgCl 2 at -20ºC is recommended. Do not store Buffer B at 4 C! Additive: Solution S (10x) may enhance the amplification of difficult DNA templates (e.g. GC-rich templates). It should only be used in defined concentrations (1x, 2x or 3x concentration) and in addition to Reaction Buffer B or BD. Note: Solution S does not replace the Reaction Buffer. It is an additive if unspecific amplification results are to be expected! 3

4 Protocol Pipette the components in the order described below: Components Reaction Buffer B or BD if necessary: Solution S Volume/ end-concentration 1x H 2 O MgCl 2 end-concentration 2,0-2,5 mm; recommended: 2,5 mm dntps 200 µm DNA template 5ng-100 ng/100 µl reaction volume Primer pmol/100 µl reaction volume Polymerase 2-5 U/100 µl reaction volume, for difficult templates up to 10 U/100 µl 4

5 Technical Data Source: Purified from an E.coli strain carrying Taq-DNA-Polymerase overproducing plasmid. The original enzyme has been isolated from Thermus aquaticus. Analysis conditions: 25 mm Tris HCl (ph 9,0 at 25 C), 50 mm KCl, 2 mm MgCl 2, 0,1mg/ml gelatine, 200 µm each datp, dgtp, dttp, 100 µm [α32-p]dctp (0,05 µci/nmol), 12,5 µg activated salmon sperm DNA. Unit definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol dntp into acid-insoluble form in 30 min 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: Taq-DNA-Polymerase is a highly processive 5 à 3 -DNA-polymerase with 5 à 3 -exonuclease activity. 3 à 5 -exonuclease activity lacks completely. Additionally, the enzyme adds nucleotides (mostly adenosines) to the 3 -ends of the DNA, so that TA-cloning is possible without further modifications. Application & Quality control: Primer extension reaction: the enzyme is free of nicking and primer extension activities as well as of exonucleases and unspecific endonucleases. SDS/PAGE: 95 kd-band, purity >98%. Activity and stability tested via PCR. The error rate per nucleotide per cycle is ~8.3x10-5 ; the accuracy ~ 1.2x10 4. Estimated half life at 95ºC: 90 min. Quantity: 500 Units / 100 Units Concentration: 5 units/µl Shipment: Storage:: at room temperature at -20 C 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, are worn. Care should be taken to avoid contact with skin or eyes. In case of contact wash immediately with water. Note: Some applications in which this product is used may require a license which is not provided by the purchase of this product. Users should obtain the license if required. 5

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7 Other products that might interest you: Universal Agarose Universal Agarose is a cost-saving standard agarose ideal for analytical and preparative separation of nucleic acids in the range of 0,05-50 kbp. 500g for only 189 EUR plus MwSt. Low Melt Agarose Low Melt Agarose is is a high-quality agarose with a relatively low melting point, ideal for the separation of nucleic acids in the range of 0,8 25 kbp. Due to its high-resolution capabilities it is suitable for all preparative gel electrophoresis. 25g for 94,90 EUR plus MwSt. High-definition Agarose High-definition agarose as an alternative to labor-intensive, expensive polyacrylamid gels. It is optimized for the separation of nucleic acids in the range of 20bp 800bp. 25g for 81,90 EUR plus MwSt. The agaroses with the best price-performance ratio: 7

8 Other products that might interest you: DNA-Markers and DNA-Ladders The Bio & SELL DNA-Markers and DNA-Ladders are characterized by sharp bands and excellent resolution in every size range. 100 bp DNA- Ladder premium including loading buffer Suitable for the size determination of double-stranded DNA in the range between 100 bp and 1,500 bp. 1 x 50ug for 46,90 EUR plus MwSt. 1 kb DNA-Ladder including loading buffer Suitable for sizing double-stranded DNA in the range between 250 bp and 10 kb. 1 x 50ug for 46,90 EUR plus MwSt. DNA-Markers: Sharp in every size range! 8