Klearkall MasterMix
General PCR KLEARtaq - Hot start Taq
TaqMan Vs KlearKall Data
Corn
Bovine
Human
Introduction
KlearKall mastermixes (MM) contain Klear Taq DNA polymerase (more info here) in a highly optimised reaction buffer, along with dNTPs required to perform both general and real-time PCR reactions. This formulation gives the user the convenience of a one liquid addition to speed up the PCR setup process along with giving the confidence that each PCR reaction will be the same.
This formulation is in daily use at KBiosciences for general PCR and end-point determination of Taqman reactions, and has been proven time and again to be robust and high quality
An example of the KlearKall MM + Rox in a
Taqman end point SNP genotyping reaction
Quality Control
Each batch is quality controlled for both performance in general PCR & Taqman end-point.
To assess general performance KlearKall MM is tested in a difficult PCR known to perform badly when non-Hot Start Taq polymerase is used (this PCR suffers badly from primer-dimer without the use of Hot Start Taq). A 850bp fragment of the VEGF gene is amplified and the mix is only passed when a clear single band is visible.
To assess end-point Taqman performance KlearKall MM is tested using an ABI Assay on demand SNP genotyping probe and primer set. The mix is only passed with a minimum of 98% call rate, no NTC movement and clear defined clusters when compared to the previous and a commercially available alternative. We have recently performed a number of in-depth analyses of the performance of Klear Kall to ABI's two Taqman mixes (the data can be found here).
Real-time PCR performance is assessed again using a dilution series of human genomic DNA and an the same Assay on demand set as above. The mix is again only passed when the standrad curve is shown to be the same as the previous batch and also in relation to a commercial alternative.
Uses
KlearKall Mastermixes can be used for many PCR applications. They are available with and without ROX passive reference dye, and are suitable for all PCR applications and are a direct replacement for all leading commercial Hot Start Taq mastermixes.
Validation, Testing & Performance
KlearKall MM's have been tested against all leading competitor enzymes and compare very favourably with all. We have used it in house for some time for the following applications, general PCR, library PCR, nested PCR, Whole genome amplification (PEP) & Taqman end-point determination.
Three different sources of genomic DNA were assessed with three different Taqman reactions mixes, two from ABI and Klear Kall.
It should be noted that the DNA was chosen due to its low quality to emphasise the difference that the Taqman mix can have on the data.
DNA extracted from Corn
DNA extracted from Corn
DNA extracted from Bovine material
DNA extracted from Bovine material
DNA extracted from Human genotyped on a mitochondrial SNP
DNA extracted from Human genotyped on a mitochondrial SNP
