For a standard procedure of DNA extraction, what will happen if you did not add the concentrated salt solution but proceed with centrifuge and subsequently removing the solution and add in alcohol?
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NemaToStella
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Posted 07 March 2010 - 01:23 PM
Are you refering to DNA precipitation? If you don't add salt, the DNA won't precipitate and hence will remain in solution and you won't get a DNA pellet.
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Denny
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Posted 07 March 2010 - 03:50 PM
PTeo, on Mar 7 2010, 03:09 PM, said:
For a standard procedure of DNA extraction, what will happen if you did not add the concentrated salt solution but proceed with centrifuge and subsequently removing the solution and add in alcohol?
You need the salt (positively charged ions) in Ethanol precipitation of nucleic acids, to bind to negatively charged phosphates exposed by the ethanol for precipitation to occur.
If your lab has a copy of "Molecular Cloning - laboratory manual," by Sambrook and Russell, check it out. Manuals like these are a great source of information about the how and why for techniques used in the lab. They give an explanation of the purpose for the reagents in your reactions and will give you a better understanding of the history, theory, and chemistry behind the protocols. The where we've been and where we're going and why perspective.
Understanding what's going on in that tube on your bench will help you to troubleshoot, optimize, and develop new techniques in the future.
