I want to avoid ethanol as it is known to hinder downstream amplification process. Also, I am in a process of developing an automated diagnosis of ZIKA and would want to avoid phenol/chloroform and other chaotropic agents which are toxic. This is because my project is aimed at developing a point of care diagnostic device for on field trials in disease endemic areas and would want to use a non-toxic reagent as my wash buffer. Until now, my results by using Nuclisens Wash Buffers as well as 70% and 91% ethanol as wash buffers have been encouraging but I feel that ethanol hinders the downstream amplification step and would want some alternative if possible. Any suggestions?
Actually for POC you may ignore the step of extraction, and lysate directly to PCR. Or you may use some commercial products and protocols for direct PCR right after lysis procedure, i.e.: prep-and-go (Thermo), AmpSolution (Promega).
You can use commercial protocols for direct PCR right after lysis step.
Thank you for your valuable answers, but I was wondering if there were any associated major disadvantages associated with commercial direct PCR products? If someone could guide me regarding this it would be really helpful.
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