FS Fast DNA Polymerase

FSTM Taq DNA polymerase is the latest generation Taq-based DNA polymerase. It possesses high amplification efficiency as does Taq polymerase, and fast elongation ability as does KOD polymerase, can be used in a variety of PCR. The FSTM PCR Buffer, designed for FSTM Taq DNA polymerase, can be used in fast amplification reaction. The elongation rate of FSTM Taq DNA polymerase is as fast as 5s/kb for simple templates, which shortens the amplification time at least by half . Contents FSTM Taq DNA Polymerase, 10x FSTM PCR Buffer, 6 x gel loading buffer Features Fast rate of elongation: elongation rate can reach to 3 kb/min, more than twice the rate by regular Taq DNA polymerase; Highly thermostable : have a half-life of over 40 min at 95°C incubation Generates 3'-dA overhangs PCR products. Applications Routine PCR PCR labeling PCR sequencing Generate PCR product for TA cloning Unit Definition One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nM of dNTPs into an acid-insoluble form in 30 minutes at 70°C using hering sperm DNA as substrate. Quality Control The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases is confirmed by appropriate quality tests Functionally tested in amplification of a single-copy gene from human genomic DNA Storage Buffer 20 mM Tris-HCl, 1 mM DTT, 0.1 mM EDTA, 100 mM KCl, 0.5 %TW 20, 0.5 % NP 40, 50 % Glycerol 10X FSTM PCR Buffer 200 mM Tris-Cl(PH 8.8), 100 mM KCl, 100mM (NH4)2SO4, 16 mM MgSO4, 1% Triton-X-100. Store all components at –20°C Protocol for PCR All solutions should be thawed on ice, gently vortexed and briefly centrifuged. Add in a thin walled PCR tube on ice: For a total 50μl reaction volume Reagent Volume (50 µl rxn) Final concentration 10x PCR Buffer 5 µl 1x dNTPs(10 mM each) 1 µl 0.2 mM each Primer I Variable 0.4-1 µM Primer II Variable 0.4-1 µM Fast DNA polymerase (5U/µl) 0.25-0.5 µl 1.25-2.5U/50 µl Water Variable to 50 µl N.A. Gently vortex the sample and briefly centrifuge to collect all drops to the bottom of the tube. ·Overlay the sample with mineral oil or add an appropriate amount of wax. This step may be omitted if the thermal cycler is equipped with a heated lid. ·Place samples in a thermocycler and start the program. PCR Cycling Protocol Initial Denaturation 94°C 3 min 25-35 Cycles 94°C 55-68°C 72°C 30s 30s 10-60 s Final Extension 72°C 2 min