Does anyone have an easy to follow protocol for the preparation of home-made taq polymerase 1? If so, which clones do you use and how can I get them? Kindly assist.
Dear Muriira,
in my experience producing Taq polymerase yourself is not neccessarily as cost-effective as people think unless you really need a ton.
It is a lot of work and does need reasonably experienced personnel to perform.
If you need a lot of polymerase, you could get a quantity quote from big suppliers (although they may not be very cheap) - or you could look at using something like alibaba.com to get it directly from producers in China. We did that for getting 50 g of statin for animal work that would have cost £2000 from Sigma - we could have gotten several kgs through Alibaba for that price!
However, your employee costs are probably low in Kenya in comparison to England, so it may be worthwhile...
have a look here
http://bitesizebio.com/10423/free-pcr-for-5-years-or-how-to-make-your-own-taq-and-pfu/
and you can get plasmids cheaply from here:
http://www.addgene.org/25712/
http://online.liebertpub.com/doi/abs/10.1089/ind.2010.6.295
This is a less than 24 hours protocol (including expression purification and buffer changes)
Thank you all of you for the wonderful insight. I need to do this in large scale and am interested in coming up with a set up to be able to supply a number of labs am working with. Krishna? I cannot download your article. Kindly if you dont mind, send a PDF to me.
Regards.
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