Neutralization Buffer B (for transfer to nylon membranes)-ML035-100ML
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Catalog No.: ML035-100ML
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Neutralization Buffer B (For transfer to nylon membranes), is supplied by HiMedia. It is available in the form of a colourless solution with a pH in the range of 7.1 to 7.3. Neutralization Buffer B is mainly used during Southern blotting procedure where DNA is transferred from agarose gel to positively charged nylon membranes. This buffer lowers the pH of DNA during the blotting process and comes as a ready to use solution and can be directly used. Neutralization Buffer B is composed of 0.5 M Tris and 1 M Sodium chloride and the pH is adjusted to 7.2. It is advisable to store this product room temperature (15-25C).
Southern blotting, first devised by E. M. Southern in 1975, is one of the prime techniques in molecular biology. This procedure involves the transfer of DNA bands, usually restriction fragments, from agarose gel to a nitrocellulose or nylon membrane. If a nitrocellulose membrane is being used then the agarose gel is treated with Neutralization Buffer A after the denaturing step. This treatment is essential as DNA does not bind to nitrocellulose at a pH of greater than 9.0 and this Tris-containing buffer brings the pH back to proper binding environment. Neutralization Buffer A is mainly used to neutralize DNA agarose gels after the DNA denaturation in base step before transfer to nitrocellulose membranes.