I am aiming to amplify a 1344 bp DNA region. Can you suggest me a good and high yielding Polymerase enzyme for the purpose? Also, the enzyme must be cost effective.
Hi Rajshree
We have been using Taq DNA polymerase from Bangalore Gene for routing amplifications of DNA fragments up 3 kb...If your prupose if only PCR based analysis (not cloning and sequencing) this is ideal and also cost effective....if you plan to use it for sequencing etc. then you can even go for a proof reading polymerase (Pfu, Pwo or a High-Fidelity Polymerase for Roche)
bye
...Taq Gold Pol!
Kit AmpliTaq Gold DNA Applied Biosystem!
Protocol: http://www3.appliedbiosystems.com/cms/groups/mcb_support/documents/generaldocuments/cms_041347.pdf
Rajshree, it can be a useful idea a longer extension step. Conventional polymerase usually extends 1,000bp per minute....Think about it and good luck.
Hi Rajshree
We have been using Taq DNA polymerase from Bangalore Gene for routing amplifications of DNA fragments up 3 kb...If your prupose if only PCR based analysis (not cloning and sequencing) this is ideal and also cost effective....if you plan to use it for sequencing etc. then you can even go for a proof reading polymerase (Pfu, Pwo or a High-Fidelity Polymerase for Roche)
bye
@alethesia souza: thanx, so can I be assured that a conventional polymerase will provide a good result if I just increase extension time?
@carlos: What is the role of BSA in increasing processivity of polymerase? never heard about that.
With any commercially available Taq DNA polymerase you can easily amplify your fragments, because it is small. For obtaining good high yield, rather than Taq DNA polymerase other parameters like initial template DNA, MgCl2, dNTPs and extension time have to be optimize. For cycle number grater than 28 or longer extension time usage use any Taq DNA polymerase stabilizing agents like BSA.
Rajshree, you can give a try. It is not possible be sure that it is gonna work. Some manufactures make sure that their Taqs amplify products bigger than 1.000bp. However, my personal experience showed that conventional PCR doesn't amplify a product so big. But I want emphasize that I never used Taq Gold nor BSA. If reagent and/or time are not problem for you, it doesn't hurt test those ideas described afore. Good luck
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