In RNA extraction we are not expecting to break the nuclear envelope.So, there is no need to use a strong lysis solution.
What is the main difference between DNA or RNA extraction?
What should the pH levels be?
RNA is very sensitive to RNAses. Guanidinium is a very strong denaturing agent so that it both dissolve cells and denaturates endogenous RNAses.
RNA is very sensitive to RNAses. Guanidinium is a very strong denaturing agent so that it both dissolve cells and denaturates endogenous RNAses.
Differences would be different, depending on what type of RNA you want to extract.
Remember RNAases are Proteins which are the most stable proteins. You need strong denaturants such as Guanidium. This will destroy the RNA degrading enzyme so that you could get good quality RNA. Whereas when you are isolating DNA you need to worry about DNA degrading enzyme (DNAase) which are relatively easy to degrade with reagent such as phenol. Best of wishes.
in my opinion,,. rna extraction is much more delicate than dna extraction because of the edlicate nature of rna. so prime difference between the procedures are that rna extraction is sensitive to any impurity or temperature much more than the dna. of all the impurities, rnases being the most hazardous ones. rnases are all around us and we consistently secrete them through our skin as well. this high abundance of rnases in the enivironment demands a high efective rnase destroyer like guanidium chloride (or guanidium thiocyanate) which destroys not just rnases but most protein by inverting their structures and adduct formations. the proteins which were folded in a aqueous environment in cell get reversed when in presence of guanidium thiocyanate. that is all i know.. hope it helps. and also you may want to break the nuclear envelope based in the rna you are looking for. extraction process changes based on the different classes of rna you are planning to extract.
Hi, I have an identical issue. while trying to isolate RNA from orchid roots I am referring a protocol where guanidine sulphate is used. But in my lab I donot have at present. Instead I have Guanidine isothiocyanate and guanidine hydrochloride. Can i supplement it with any one of them?
RNA lysis buffers that contain guanidinium thiocyanate or guanidinium–HCl reproducibly yield very high-quality RNA samples. This is true because of the extremely chaotropic nature that these chemicals exhibit; they are among the most effective protein denaturants (Cox, 1968; Nozaki and Tanford, 1970; Gordon, 1972)
https://www.sciencedirect.com/science/article/pii/B9780123747273000024
RNA Isolation Strategies
Robert E. Farrell Ph.D, in RNA Methodologies (Fourth Edition), 2010
I need to do whole genome DNA extraction from pellets of deer family. The tissues are quite hard. I tried various conventional methods for DNA extraction, but the results were not good. However, I found one method in which lysis buffer Gu-thiocyanate is used, but I do not have it my lab. Can I use GuHCL instead for lysis buffer.
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