Taq DNA Polymerase is the workhorse driving DNA amplification in Polymerase Chain Reactions (PCR).
Gene And Cell Technologies Taq DNA Polymerase is produced in a large scale bioreactor process, followed by multiple orthogonal HPLC chemistries. The result is one of the cleanest enzymes you will find on the market.
Taq DNA Polymerase should be used at 1 - 10 units per 50 ul reaction. In general, longer templates will require less polymerase than shorter templates. For detailed PCR optimization protocols, please refer to widely published PCR guidelines. This enzyme will generally behave as expected according to standard PCR protocols.
Please note that this bulk Taq Polymerase preparation does not include buffers. The intention with the present product is to give you the most enzyme for the least amount of money. For a more convenient, ready-to-go version, please refer to our 5x PCR MasterMix.
Pipetting Plan for Standard PCR
Water ........................... 31 ul
Buffer (10x)*.................. 5 ul
dNTP (10 mM)*............. 1 ul
Primer F (10 uM) .......... 1 ul
Primer R (10 uM) .......... 1 ul
Template DNA ............... 1 ul
Taq DNA Polymerase....0.25 ul (1.25U)
Some of the water volume may be replaced with other reagents as needed. This could be an increased amount of template (assuming it is clean and free of PCR inhibitors), or a dye or probe for qPCR. The entire reaction can be scaled to different volumes, e.g. 50 ul, by multiplying all the volumes accordingly.
In the experiment shown below, we amplified a test plasmid by qPCR. We varied the amount of Taq DNA polymerase, to show the effect on the qPCR chart and gel bands. This experiment is routinely used to determine the activity of our new production batches.
As can be seen, there is an optimal amount of Taq DNA Polymerase in qPCR, that gives a steep fluorescence curve, in combination with a clean gel band. In the case of this 20 ul qPCR reaction, it is in the range of 0.7 - 0.8 units. Higher amounts of polymerase result in slight non-specific amplification (hints of an unwanted band are visible at the three highest concentrations). Lower amounts of polymerase result in increasingly flatter curves, less suitable for quantitative applications. However, if ultimate sensitivity is not required, and the budget is limiting, substantially lower amounts of Taq than recommended can result in good amplification at later cycles.
This experiment should be viewed as a starting point for your own studies. When ultimate sensitivity is the goal, each new qPCR reaction should be optimized individually. This experiment shows that the amount of Taq DNA Polymerase in the reaction is one important variable to consider.
Shipping & Storage
Taq Polymerase is stable over a wide range of temperatures and ships at ambient temperature. This procedure has undergone extensive testing. We shipped this polymerase across the country and back multiple times, and found no adverse effects on PCR activity, in qPCR and regular reactions.
Upon receipt, it can be stored in the fridge for at least 1 year, or in the freezer at -20C or -80C for at least 5 years. If it is ever left at room temperature by accident, there will be no adverse effects for many days.
Unrestricted Use License
It's OK to use Taq DNA Polymerase from Gene and Cell Technologies for your own research, for the manufacture of commercial products, direct re-sale, or any other legal purpose. Purchase of this product does not grant a license to any patent. This product has not passed any kind of regulatory muster, and is not presently approved for use in any diagnostic or therapeutic procedures.
We use Gene And Cell Technologies as our supplier at Berkeley Biolabs, the largest biotech hackerspace in the world. I can assure you that Gene And Cell Technologies ultrapure Taq Polymerase work just as well as the expensive stuff from the big companies. There are no false positives with bacterial or M13 primers, and the bands are incredibly strong. Thanks Gene And Cell Technologies for helping make our biotech hackerspace possible with your very startup-friendly pricing and high quality products.
Ryan Bethencourt, Founder at Berkeley Biolabs.