search for plasmid containing Taq gene

Cathal Garvey via methods%40net.bio.net (by cathalgarvey from gmail.com)
Wed Jul 28 02:15:54 EST 2010


@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" <biocjh from 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 <dk from no.email.thankstospam.net>:

> In article <mailman.912.1280230802.25217.methods from net.bio.net>, jh <
biocjh from gmail.com> wrote:
>>Dear...


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