Q: I want to amplify a long fragment with high fidelity, what enzyme should I use?
A: You should use LA version of any DNA polymerase listed in our website (Click here).
Q: I get false negative result even after DNA purification, what is the reason and how to solve this problem?
A: Your DNA sample may contain some residues of PCR inhibitors that lead to false negative result. You can use our inhibitor-resistant mutant enzymes in the PCR (Click 1，2). Furthermore, you can use both the inhibitor-resistant enzymes and PCR enhancer cocktail (PEC) in the PCR. You can find the table in “PCR Enhancers” category in the website from which you know what PEC is suitable for your sample (Click here).
Q: I want to directly amplify DNA gene target from blood or blood sections, what product should I select?
A: We suggest that you select OmniTaq or KlenTaq combined with PEC in PCR if the concentration of blood in PCR is not too high or select our Direct Blood PCR Kit (Click here) or Direct S/P PCR Kit (Click here). We have a new generation of Taq and Klentaq mutants, which are able to tolerate even higher level of PCR inhibitors (Click 1, 2).
Q: I want to directly amplify RNA gene target from blood or blood sections, what product should I select?
A: You can select our Direct Blood RT-PCR or Direct S/P RT-PCR Kit. For more information, click here.
Q: I want to screen bunch of colonies, what product should I use?
A: Our Direct C-Screening PCR Kit is a good choice, instead of growing colonies and plasmid purifications (Click here).
Q: Do I have to use PEC in PCR?
A: If you use purified DNA as a template, we don’t suggest PEC in the reactions because it may inhibit PCR. However, if you want to amplify the gene target directly from the crude samples, PEC is able to facilitate the performance of the mutant enzymes.
Q: Which PCR enhancer should I select?
A: Different PEC may facilitate the performance of Taq mutant enzymes in different crude samples. Please refer to the Table under product category "PCR Enhancer" or "Selection of PCR enhancers" (Click here) for the details.
Q: I am designing a qPCR, what enzyme should I select?
A: If you do qPCR with SYBR Green or Eva Green, you can use any of our mutant enzymes. However, if you design the experiment with TaqMan assay, you should select plain Taq or our full-length Taq mutants (Click here). This is because the Klentaq family is a truncated version that lacks 5’ to 3’ exonuclease activity.
Q: What is the unique feature of your Direct PCR / RT-PCR Kit?
In most case you can directly amplify the gene target from crude samples without lysis step which is required in other similar products (Click here).
Our novel Direct One-Step RT-PCR Kits are unique. With these kits, you can directly amplify RNA gene target from serum, plasma, whole blood and other body fluids without RNA purification (Click here).
Q: What is difference between your hot-start enzymes and other commercial hot-start enzymes?
A: Our hot-start enzymes not only have hot-start feature but also tolerate high concentration of PCR inhibitors (Click here)
Q: Almost all companies ship DNA polymerases with ice pack or dry ice, why do your company ship at ambient temperature?
A: Taq mutant enzymes are very thermostable. The mimic experiment, in which the enzymes were exposed to field for 30 days during summer time, showed the enzymes didn't significantly compromise functionality under this stringent condition (See data). We ship the enzymes worldwide for many years and there isn't any problem.
Q: I want to detect rare mutation in the presence of large excess wild type allele, what enzyme should I use?
A: You should select KlenTaq-P (Click here). KlenTaq-P is a 5’-exonuclease deficient Taq polymerase (an N-terminal deletion of Taq) with improved fidelity and thermostability, plus an extra mutation rendering the enzyme PAP function (Pyrophosphorolysis-activated polymerization). The enzyme is capable of detecting rare mutations, large heterozygous deletions, gene duplications, etc. in the presence of a large excess of wild type allele and inorganic pyrophosphate). In theory PAP can detect a single base mutation in 3x1011copies of wild type allele. KlenTaq-P LA is a blending of KlenTaq-P and a proofreading enzyme.
Q: May I use KlenTaq-P in TaqMan assay?
A: KlenTaq-P is a 5’-exonuclease deficient Taq polymerase, so it alone can not be used in TaqMan assay. However, you can blend it with full-length Taq or Taq mutant enzymes to achieve this aim (Click here).
Q: How many units / ml of your enzymes?
A: We prefer to provide the enzyme in reactions / volume format, rather than Units / µl concentration. The reason is that the Taq DNA polymerase units are not defined in a real PCR assay, but in "nucleotide incorporation assay", and sometime it may not predict the enzyme performance in PCR amplification. Our better alternative is that we measure the functional concentration of the enzyme in a two-round PCR enzyme titration test, to calibrate and normalize the enzyme PCR strength to our "1X strength" for each lot. We believe this approach is more practical and useful for the final users, as they'll know how many PCR reactions they can perform with defined amount of enzyme based on the target size. You can find the details in Data Sheet of all enzymes.