Hello,
I would like to extract DNA from dried insect museum samples and try to amplify a short mitochondrial gene fragment (short portion of cox1). Reading several articles on this topic, I notice that it is quite common to use high specificity Taq enzyme. However, appears not to be convergence on a specific product or type of enzyme. So I would like to ask for advice, based on your experience, on what might be the best Taq DNA Polymerase and/or best practice for trying to amplify cox1 short fragment from low concentrations and degraded DNA from museum sample extraction (in particular insects).
Just for knowledge, the products that I have seen several times in the newspapers are:
- Platinum™ Taq DNA Polymerase Catalog number: 10966018,
- AmpliTaq Gold™ DNA Polymerase with Buffer II and MgCl2 Catalog number: N8080241
- HotStarTaq DNA Polymerase (250 U) Catalog number: 203203
- TAQKB Roche KAPA Taq PCR KitCatalog number: KK1014
Hi Emmanuele,
The reason there is no one recommended enzyme is that there are LOTS that work really well. I'd advise a "hot start" enzyme since that will reduce primer artifacts. I'm also a fan of the ready-to-use PCR mixes (enzyme, buffer, MgCl2, etc. all in one tube). Makes it much easier to have consistent results.
You should have a really robust positive control, test out your primers and cycle conditions on non-precious samples first, and always include a full set of controls.
Good luck!
A particularly good one is:
QIAGEN Multiplex PCR Kit
hot start, master mix, extremely specific and used in forensic laboratories.
Thanks Filipe for the suggestion, I will consider this Qiagen Kit!
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