@DK- Have you successfully gotten restriction enzymes too then? Because those patents occasionally mention having to transform the bacteria with the methylase first so it has time to work. Perhaps growing cells at a lower temperature for a while could help slow the cutting enzyme, but perhaps this would slow the methylase equally? I wouldn't be concerned about this for sending my own proteins, as I favour making coding sequences available for my work anyway. Life is too short to delay others' work for the sake of exclusivity! :)
@C J- Back on the topic of taq.. it depends. You could try primers for the wild-type sequence found in Thermus aquaticus, but it's equally possible these days that the coding sequence is codon optimised into something entirely new. If the wild type won't amplify, you could try making primers for an e.coli optimised sequence (Google 'jcat codon', but it'll be hit and miss. You could improve matters by making degenerate primers to account for common third-base variations among optimal codons, but at that rate it's turning into a whole research project of its own! My suggestion: either get the peptide sequence of taq from uniprot and order a codon optimised gene from Mrgene.com or one of the many equally-priced companies, or transform with taq prep like DK said and then try to sequence any plasmids that arise. --- Twitter: @onetruecathal Sent from my beloved Android phone. On 28 Jul 2010 04:16, "jh" <bio...@gmail.com> wrote: thanks a lot. but if i want to amplify the sequence, how do i design primers. do all Taq DNA polymerases have the same sequence？ CJ 2010/7/27 DK <d...@no.email.thankstospam.net>: > In article <mailman.912.1280230802.25217.meth...@net.bio.net>, jh < bio...@gmail.com> wrote: >>Dear... _______________________________________________ Methods mailing list Methods@net.bio.net http://www.bio.net/biomail/listinfo/methods
