Certain sources of DNA should not be used, such as: Your family pet, Fido the dog Your little sister's big toe Bugs you caught in the yard Step 1: The blender separates the pea cells from each other, so you now have a really thin pea-cell soup. Soapy Peas Pour your thin pea-cell soup through a strainer into another container like a measuring cup.
The basic procedure is that salt and ethanol are added to the aqueous solution, which forces the precipitation of nucleic acids out of solution.
After precipitation the nucleic acids can then be separated from the rest of the solution by centrifugation. So how does this work? Ethanol precipitation of nucleic acids is all about solubility… First we need to know why nucleic acids are soluble in water.
Water is a polar molecule — it has a partial negative charge near the oxygen atom due the unshared pairs of electrons, and partial positive charges near the hydrogen atoms see the diagram on the right. Because of these charges, polar molecules, like DNA or RNA, can interact electrostatically with the water molecules, allowing them to easily dissolve in water.
Nucleic acids are hydrophilic due to the negatively charged phosphate PO3- groups along the sugar phosphate backbone. The role of the salt in ethanol precipitation… Ok, so back to the protocol.
The role of the salt in the protocol is to neutralize the charges on the sugar phosphate backbone. A commonly used salt is sodium acetate. The positively charged sodium ions neutralize the negative charge on the PO3- groups on the nucleic acids, making the molecule far less hydrophilic, and therefore much less soluble in water.
A few tips on ethanol precipitation… Choice of salt Use Sodium acetate 0. Use Lithium Chloride 0. This is because 2. In these cases, use NaAc. Use Ammonium acetate 2M final conc for the removal of dNTPs, but do not use for preparation of DNA for T4 polynucleotide kinase reactions as ammonium ions inhibit the enzyme.
To increase the yield in precipitations of low concentration or small nucleic acid pieces less than nucleotides Add MgCl2 to a final concentration of 0. If you want to learn more about the ins and outs of ethanol precipitation and other DNA cleanup approaches, you might want to check out these… More articles on Ethanol precipitation.Extraction of DNA from Calf or Hog Thymus/Isolation of Yeast RNA I.
Abstract Nucleic acids may be divided into two groups RNA and DNA. DNA contains almost all the genetic information while RNA serves as the bridge between the DNA and proteins.4/4(1).
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Extraction of DNA from Calf or Hog Thymus/Isolation of Yeast RNA I. Abstract Nucleic acids may be divided into two groups RNA and DNA. DNA contains almost all the genetic information while RNA serves as the bridge between the DNA and proteins. 1. Publication of high quality, high impact, peer reviewed research papers..
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DNA appears fluffy which means it has sheared in the extraction process DNA appears as thin threads. Although DNA that strands is the most impressive, DNA that has sheared still shows that DNA is present.