Re: [DIYbio] polymerase expr/purification WAS:Anyone need some synthesis?

openbiotech.org or something had a kit for that I believe. master
mixes from NEB are so good and cheap though

On Fri, Mar 21, 2014 at 6:39 PM, Jeswin <phillyj101@gmail.com> wrote:
> On Fri, Mar 21, 2014 at 5:25 PM, Koeng <koeng101@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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