单位定义 在标准的 Taq DNA 聚合酶反应条件下的二核苷酸重复位点扩增中,一个单位的 Platinum™ GenoType Tsp DNA 聚合酶被确定为在功能上与一个单位的 Taq DNA 聚合酶等效。一单位 Tsp DNA 聚合酶近似2.5活性单位。在优化的反应条件下(74°C 时),一个活性单位会在30分钟内将 10 nmol 脱氧核苷酸掺入酸沉淀材料中。
What is the difference between Platinum technology and AccuPrime technology?
With Platinum technology, anti-DNA polymerase antibodies bind to the enzyme until the denaturing step at 94 degrees C, when the antibodies degrade. The polymerase is now active and primer extension can occur. AccuPrime Taq combines Platinum Taq (Taq + Platinum antibodies) with proprietary thermostable AccuPrime accessory proteins. The 10X reaction buffer contains the accessory proteins which enhance specific primer-template hybridization during each cycle of PCR.
Is there anything to prevent AmpliTaq Gold DNA polymerase from extending from the 3’ end of a TaqMan probe in a 5’ nuclease assay?
Yes. There is a phosphate group on the 3' end of all TaqMan probes that prevents such extension.
How does AmpliTaq Gold DNA Polymerase differ from AmpliTaq DNA Polymerase?
AmpliTaq Gold DNA Polymerase is a modified form of AmpliTaq DNA Polymerase that contains a proprietary chemical (or so-called hot start molecule) bound to the enzyme's active site. In order to activate the AmpliTaq Gold DNA Polymerase fully, we recommend an initial activation step of 95 degrees C for 10 min when using GeneAmp 10X PCR Buffer I and/or GeneAmp 10X PCR Buffer II and Mg in one of our thermal cyclers. When using GeneAmp 10X PCR Gold Buffer, activation time can be reduced to 5 minutes. Once activation is complete, you can proceed with your standard PCR cycling program (denaturing, annealing, extension, etc).
Does AmpliTaq Gold DNA Polymerase contain exonuclease (proofreading) activity?
No, AmpliTaq Gold DNA polymerase does not contain proofreading activity, however fidelity in PCR amplifications utilizing this enzyme may be improved. High fidelity can be achieved by: 1. Decreasing the final concentration of each nucleotide to 40-50 uM. 2. Using the lowest MgCl2 concentration possible. 3. Using less enzyme. 4. Decreasing extension times. 5. Using the highest annealing temperature possible. 6. Using as few cycles as possible.