Hi
Recently we are working with Wako DNA extractor kit and it uses wash buffer A and B to wash the precipitated DNA pellet.
Does anybody have explanation about these wash buffers composition?
This paper might help. ISOLATION AND PURIFICATION OF GENOMIC DNA
www.envfor.nic.in/.../16%20Dr%20Randhawa_%20Isolation%20%26%20purification...
by GJ Randhawa - Cited by 1 - Related articlesThe extraction buffer also contains a reducing agent (β- mercaptoethanol) and a chelating agent ...DNA can be precipitated using either ethanol or isopropanol. ..... DNA quantification is an important step to know the amount of isolated DNA. ... A good DNA preparation exhibits the following spectral properties: A300< 0.1.
Usually when you do DNA extraction, the first step is a chaotropic salt like guanidine thiocyanate or sodium iodide. After, you need to do a salting out process to remove all added salts that could inhibit PCR or any other process carried out later. In this case, following buffers will contain more water and less salts to wash your preparation. Several DNA extraction kits use this methodology.
Dear apply chaotropic salt and salting out process by which the DNA could be washed.
Hi Hossein Behrouz ,
As you have figured out from the previous answers, i would like to add that the Wash Buffer A (used first) generally comprises of 3M sodium acetate in 75% Ethanol solution and the Wash Buffer B (used lastly) has a higher percentage of Ethanol (~90%).
Hope it helps,
Subham.
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