Does anyone have a / access to a proofreading polymersase?

[Mega]

Hi everyone, 

I soon need to do a PCR of a ~10 kbp fragment. 

I asked in every lab of my university if they had some proofreading polymerase (like Pfu). Because the error rate of Taq polymerase is somewhat like 1:9000, it won't work with > 3 kbp. 

Buying one myself may not be an option, because I was told that those are remarkably expensive... (Althought, here it seems quite affordable... http://www.thermoscientificbio.com/pcr-enzymes-master-mixes-and-reagents/pfu-dna-polymerase/ 60€ for 100 units )

Of course, I'd pay the mailing fee, and a little compensation. Or tade it for one of my plasmids. pVIB, pGreenII-0049, p35S-GFP. Have acces to pTracer-CMV2, so I could possibly share also this one. 

Just shoot me a mail if interested ;) 

[Mega]

Just read this... 

Formulation: Genscript Taq DNA Polymerase has been formulated using a proprietary technology, and the enzyme can be shipped at room temperature or stand at 37ˇăC for 7 days without losing any activity.  

Does anyone know is that is true also for pr-polymerases that they may be shipped at room temperature? If so I kan kindly ask at the company when ordering primers to include a few units as a sample ;)

[poli]

Call NEB and ask for a free sample of Q5 https://www.neb.com/products/polymerases-and-amplification-technologies/pcr-polymerases/q5-high-fidelity-dna-polymerases/detail. My lab has gotten about 5 free samples though we can get phusion for $127 per 200 rxn.

[Nathan McCorkle]

NEB has dished out free samples to me too, when I get $$$ for some
serious research I'll definitely buy from them. I also recommend them
to lots of folks because of this reason, so if you're at a big
University or company, it might be cheap advertising for them.

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--
-Nathan

[Andreas Sturm]

You think they would send it to my university for free?? Even shipping included?

Another company in Europe told me that shipping polymerase has to be on wet ice, costing additional 19€.

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[Nathan McCorkle]

It would probably be better to find a company closer to your country. I've always received samples completely free of any charge. Asking to pay shipping would help your case if they don't wanna ship to you. I've always received cold stuff in Styrofoam boxes packed with dry ice.

[Josiah Zayner]

An easy solution would be to create multiple sets of primers and amplify regions and then stitch them together or do some overlapping extension PCR. This should be a viable option using Taq and splitting the DNA into 3-4 fragments.