Bst DNA Polymerase - Large Fragment
Bst DNA allows sequencing through problematic secondary structures
Features:
- reverse transcription activity
- increased activity
Applications:
- nucleic acid amplification methods, including isothermal amplification
- whole genome amplification
Description:
Bst DNA Polymerase, Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the 5´ → 3´ polymerase activity, but lacks 5´ →3´ exonuclease activity.
Enzyme provide strong strand displacement activity. The optimum temperature is from 60-65 ⁰C. Bst becomes heat inactivated at 80 ⁰C. Bst Polymerase, Large Fragment is prepared from an E. coli strain containing a gene of the Bacillus stearothermophilus DNA Polymerase gene, lacking the 5´ → 3´ exonuclease domain.
Storage condition: @ -20°C in 10 mM Tris-HCl (pH 8.0 @ 25°С), 10 mM KCl, 1% BSA, 0,02% Tween 20, 50% glycerol.
Unit definition: One unit is defined at the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.
Purity: > 97 %
Heat inactivation: @ 80°C for 20 minutes.
Concentration: 8 U / µl
Reaction buffer (10X): provided
MgSO4: Provided in a separate Tube
Storage: -20°C
Transport: on blue ice
Note:
- Recommended for long term storage: 0.1% Triton X-100
- Reaction temperatures above 70°C are not recommended.
- Bst DNA Polymerase cannot be used for thermal cycle sequencing.
References / Protocols / Notes / Recomendations
Typical LAMP Protocol with Bst DNA Polymerase
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Bst DNA Polymerase:
Applications:
· LAMP
· Reverse transcription
· Whole Genome Amplification
Final price excl. shipping costs3
- verfügbar / avaílable
- 1 - 3 days for delivery / 1 - 3 Tage Lieferzeit1
1. A LAMP Primer Mix can be prepared with all 4 or 6 (with Loop) primers. A 10X Primer Mix should contain: 16 µM FIP, 16 µM BIP, 2 µM F3, 2 µM B3, 8 µM LoopF, 8 µM LoopB in TE or water
2. Reactions should be setup on ice.
3. If analyzing via agarose gel electrophoresis or other method requiring opening LAMP reaction vessels, setup secondary analysis area and equipment to avoid contamination.
4. Running a no-template control is strongly recommended to ensure amplification specificity.
5. If optimization is desired, try titrating Mg2+ (4–10 mM final) or Bst DNA Polymerase, Large Fragment (0.01–0.2 U/µl), or changing reaction temperature (50–68°C).
6. dNTP should be used with dTTP, not dUTP
7. We recommend using LAMP primer design software such as Primer Explorer (http://primerexplorer.jp/e/).
Deutsche Beschreibung
Bst DNA Polymerase
Datasheet Bst DNA Polymerase (large fragment)
Material Safety Datasheet
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GeneOn GmbH
Paracelsusstrasse 10
67071 Ludwigshafen am Rhein
Deutschland / Germany
Phone: +49-621-5720 864
Fax: +49-621-5724 462
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