dex.z

thanks for the responses. But im still not entirely sure how vacuum baking causes the dna to be fixed to the nylon membrane. Im aware of other methods (such as UV crosslinking) but for my particular case I have to explain how it happens in terms of heating the membrane to 80C for 30 mins. From this site: http://www.invitrogen.com/site/us/en/home/References/Ambion-Tech-Support/northern-analysis/tech-notes/membrane-transfer-and-crosslinking-for-rna.html They give the following explanation: BakingBaking works by heating the membrane to drive out all water solubilizing the RNA. A large component of RNA is its hydrophobic nucleotide bases, which make hydrophobic contacts with aromatic groups on the membrane. This interaction is affected by heating in an oven at 80°C for 15 min. The only danger in baking is that the membrane can be damaged if the heat is not regulated to prevent temperatures from rising much higher than 100°C. Is it safe to assume single stranded DNA undergoes similar process? Since the only difference between RNA and single stranded DNA is in the ribose which doesn't seem to have an importance in its fixation to the membrane..

This is my first post. so apologies for breaking any posting rules. My question regards a pre hybridization procedure in which the denatured DNA is transferred onto a nylon membrane. To do this the nylon membrane is baked which allows the DNA to covalently bind to the membrane, through a dehydration reaction. Or so I was told by my university tutor. However, I'm not sure of the chemistry behind this reaction. If anyone could explain or reference a source that can explains the chemistry concisely I would appreciate it. Thanks.