SilverStar DNA Polymerase

Details for Product No. ABIN3188323, Supplier: Log in to see
Application
Polymerase Chain Reaction (PCR), DNA Amplification (DNA Amp)
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Purpose SilverStar is a high-performance thermostable recombinant DNA polymerase, isolated from Thermus aquaticus.
Specificity SilverStar is a high-performance thermostable recombinant DNA polymerase, isolated from Thermus aquaticus. It is an extremely robust Taq DNA polymerase that does not require optimization and gives high yields with minimal background consistently from one reaction to another. It is a highly active enzyme that produces excellent results across a wide range of applications such as DNA Array.
Characteristics Quality Control:
  • Each lot is tested for the absence of nicking and priming activities, exonucleases and non-specific endonucleases.
  • It is specially purified so as to remove traces of DNA that could interfere with some reactions and is free from all non-specific endonucleases, nickases and exonucleases.
Features:
  • High yields
  • Robust PCR
  • Highly purified Taq polymerase
  • No optimization needed
Components Components:
  • SilverStar DNA polymerase (100μl)
  • 10x reaction buffer (yellow cap) (2x 1.5 ml)
  • 50mM MgCl2 (2x 1.5 ml)
Biological Activity Comment The SilverStar has a 5' ' 3' exonuclease activity and also the "extendase" activity allowing TA cloning. The enzyme lacks the proofreading activity 3' ' 5' exonuclease.

Endonuclease and exonuclease activities were not detectable after 2 and 1-hour incubations, respectively, of 1 μg lambda DNA and 0.22 μg of EcoRI-digested lambda DNA at 72°C in the presence of 15-20 units of SilverStar DNA polymerase.
Unit Definition One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form in 30 minutes at 72°C.
Application Notes The SilverStar will efficiently amplify products up to 10 kb giving high yields and optimal results for application such as array PCR product constructions.

MgCl2 concentration: this polymerase is a magnesium-dependent enzyme. The supplied 50 mM MgCl2 solution should be used to adjust magnesium ion concentration. Excessive amount of Mg2+ stabilizes the DNA double strand and consequently prevents complete denaturation of DNA, which reduces the extension yield. It may also stabilize spurious primer/template annealing, thus decreasing specificity.
Comment

One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form in 30 minutes at 72°C.
Protocol For a 50 μL volume:
  • 10x Reaction Buffer (5 μL)
  • 50 mM MgCl2 Solution (1.5 - 4.0 μL)
  • dNTP final concentration (250 - 500 μM each dNTP)
  • >20 mM dNTP Mix (2.5 - 5 μL)
  • Template and Primers (as required)
  • Enzyme (0.5 - 1.0 μL)
  • Water up to 50 μL
Troubleshooting and Cycle Conditions are given in the delivered protocol.
Restrictions For Research Use only
Storage -20 °C
Storage Comment SilverStar DNA Polymerase can be stored at -20°C, in a constant temperature freezer for 12 months (1). SilverStar will remain stable if stored as specified. Repeated freeze-thaw cycles will affect the stability of Reaction Buffer. The Buffer will remain stable at +4°C for a minimum of one month.