Can I use Hot Start Taq polimerase for any target? - (Jan/06/2011 )
My supervisor asked me to order polimerase with high fidelity because she will be doing some experiment with molecular genotyping of some bacterial strains, so I was thinking of ordering a Hot Start Taq polimerase, but while reading additional info, I came across a picture where it says that it improves amplification of targets that require hot start. Now, I am confused, can I use it for amplification of any kind of target gene, or only "special"? And what is so special about this genes- they higher GC-content (so it is needed a higher temperature to disrupt the strands), or?
Thank you in advance! 
Do not worry about the statement "improves amplification of targets that require hot start". All that means is that because you need a hot start step to activate the Taq enzyme you are likely also helping denature DNA that has strong stability, like high GC-content DNA. In other words, going through a hot start step (3 to 15 minutes at around 94oC to activate your Taq, depending on the brand of Taq you get) creates an environment (high temperature for a few minutes) that helps in the amplification of high-stability DNAs.
Your hot start Taq will work just fine with most templates; it just so happens that a hot start step also leads to DNA denaturation of otherwise hard to amplify DNAs.
ivanbio on Thu Jan 6 22:22:32 2011 said:
Do not worry about the statement "improves amplification of targets that require hot start". All that means is that because you need a hot start step to activate the Taq enzyme you are likely also helping denature DNA that has strong stability, like high GC-content DNA. In other words, going through a hot start step (3 to 15 minutes at around 94oC to activate your Taq, depending on the brand of Taq you get) creates an environment (high temperature for a few minutes) that helps in the amplification of high-stability DNAs.
Your hot start Taq will work just fine with most templates; it just so happens that a hot start step also leads to DNA denaturation of otherwise hard to amplify DNAs.
thank you
Hello,
Have you ever search information about pfu DNA polymerase? As far as I know it polymerizes DNA more accurately than Taq polymerase.
hi, just for sharing:
high fidelity polymerase is not the same with hot start polymerase...
She might mean pfu polymerase which have proof reading capabilities.
high fidelity taq usually is a blend of both pfu and taq together like:
high fidelity taq: Platinum® Taq DNA Polymerase High Fidelity
high fidelity taq with hot start: AccuPrime™ Taq DNA Polymerase High Fidelity
make sure which one suits your application the best.
just my 2 cents.
Eventually, when I saw all the enhancement (yield, sensitivity and specificity) features that are gained by hot start polymerase, my mind thought it also has high fidelity.
So to make it clear: hot start polymerase is suitable for small yield templates with/out high GC-content and when we have primer-dimer issues, and Pfu is best for high fidelity, because it has the proof-reading activity.
Thank you
There also are some polymerases with both hot start and high fidelity!