It was just my idea to do pfu because look at how cheap they can make taq... it would be essentially the same protocol
-- and NEB's stuff is like a dollar a PCR
On Friday, March 21, 2014 3:39:09 PM UTC-7, phillyj wrote:
On Friday, March 21, 2014 3:39:09 PM UTC-7, phillyj wrote:
On Fri, Mar 21, 2014 at 5:25 PM, Koeng <koen...@gmail.com> wrote:
>
> Hey
>
> I am doing a large synthesis order soon from gen9. However, they insist upon synthesis of at least 25kbs, and I unfortunately only need 20kbps synthesized. If anyone wants to get some genes for cheap, contact me. It is 20 cents a base pair ( .5 to 3kbp) and as a little incentive I will also include a (native) pfu polymerase gene for expression in E coli :)
>
How hard is it to express and purify polymerase in E coli? I was
browsing Engelke's paper on it. It looks complicated and not easily
done unless you got some specific equipment. Otherwise, it's very
doable. Anyone here tried this?
Engelke, David R., et al. "Purification of Thermus aquaticus DNA
polymerase expressed in Escherichia coli." Analytical Biochemistry
191.2 (1990): 396-400.
Available at: http://deepblue.lib.umich.edu/bitstream/handle/2027.42/ 28292/0000046.pdf
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