Phenol/chloroform extraction is a common technique used to separate and purify DNA, RNA, and protein from a biological specimen, such as a virus preparation. It involves the differential partitioning of DNA/pro
phenolThis chapter describes, in detail, how to extract RNA and DNA from tissue and cell culture samples using phenol:chloroform extraction, commonly referred to by the trade names TRIzol; and TRI Reagent;. Most protocol steps have additional comments from the author regarding how to proceed at ...
extraction buffer produces a highly viscous solution. The viscosity is usually due to genomic DNA. This normally has no effect on the RNA isolation (except for dictating longer periods of spinning at the phenol-chloroform extraction steps), unless the amount of dissolved tissue was indeed too ...
Adding more lysis solution and/or re-extracting with phenol:chloroform:IAA will help alleviate this problem. Also, multiple phenol:chloroform:IAA extractions can be performed to ensure the partitioning of DNA into the organic phase during the acid phenol extractions ...
Human RNA Extraction PROTOCOL(RNA提取方法) REAGENTS Guanadinium Thiocyanate 1.0 M Sodium Citrate pH 7.010% Sarcosyl2-mercaptoethanolwater saturated phenol pH 4.0-7.0 Na2EDTAMOPS (free REAGENTS >Guanadinium Thiocyanate> 1.0 M Sodium Citrate pH 7.0...
The most widely used method for high throughput RNA isolation uses glass fiber filter plates where RNA binds to the filter in the presence of high salt and alcohol concentrations, avoiding phenol/chloroform extraction that is not amendable to automation. We developed the filter pla...
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Additionally, we did not test an RNA extraction protocol with phenol-based solution alone, but only as combined with silica-membrane columns. The latter was due to our previous experience that without additional on-columns purification, the RNA purity values tend to be poor (unpublished ...
Several approaches utilize the ’single-step’ method of total RNA isolation using acid guanidinium thiocyanate-phenol-chloroform extraction, described in 1987 by Chomczynski and Sacchi [16]. Guanidinium thiocyanate lyses the cells and inactivates RNA-digesting enzymes in one step. After a ...