Академический Документы
Профессиональный Документы
Культура Документы
Home About us DNA Extraction Cytogenetics DNA PCR technology Our Products
glpbio.com OPEN
“DNA precipitation is a process in which the nucleic acid is precipitated using alcohol and salt.
Ethanol and isopropanol are the two most common types of alcohol used in the DNA precipitation
protocol.”
DNA precipitation is an important step in the DNA extraction protocols, we can say especially, for
traditional DNA extraction protocols.
DNA extraction protocols nowadays are robust and extraordinary uses such chemistry that directly
dissolve DNA.
• Precipitation of DNA
• Washing of DNA
• Dissolving DNA
A brief outline of the DNA extraction steps.
We had covered enough article on DNA extraction. Here we will directly discuss the “DNA precipitation”
step.
Key Topics:
What is DNA precipitation?
Mechanism of DNA precipitation:
Chemicals used in the DNA precipitation:
Alcohol used in the DNA precipitation:
A common DNA precipitation protocol:
Conclusion:
Share this:
“The precipitation is a process in which the reaction between solute and solvent creates insoluble
aggregate called precipitate and the process is called precipitation.”
The DNA is hydrophilic in nature which dissolves in water but not in the alcohol.
The water has a partial positive charge and a partial negative charge. The positive charge of the water
molecule interact with the negative charge of the DNA (the PO3-) and dissolve it. However, the
interaction is not so strong.
The image shows the intermolecular interaction between water and DNA.
The salt and alcohol make the DNA more water hydrophobic, once we add the salt into the DNA, the
negative charge of the DNA interact with the positive charge of the salt, instead of the positive charge
of the water.
Read our amazing article on DNA: DNA story: The structure and function of DNA
It might bind to the water but thanks to the alcohol, the “lower dielectric constant alcohol” protect the
The DNA is visible like a cotton thread and aggregate as a precipitate due to this chemical reaction.
For a more in-depth explanation on the mechanism of the DNA precipitation please read our previous
Furthermore, instead of the negative charge of the DNA, the water remains busy in interaction with the
negative charge of the alcohol that also increase the precipitation efficiency and the overall yield of the
DNA precipitate.
The entire mechanism of precipitation is based on the polarity of the solution and the dielectric
constant of the solvent.
The DNA is a polar solution and so the water is. Therefore, by intermolecular interaction, the DNA
(more preciously the nucleic acid) remains dissolved in the water.
Gsafe Red plus - Gsafe Red plus
Ad
(polar solution dissolve in polar and non-polar solution dissolve in non-polar solution)
Here, the interaction affinity is strong between the H+ and P03– of the water and DNA, respectively,
hence both interact with each other.
The entire solution contains not only the DNA but some of the other impurities too. Thanks to this, it is
necessary to separate DNA even into the next level of structure that visibly separable.
A precipitate is a visible form of DNA (Although it does not look like the spiral DNA, it is just a clump of
white aggregates), by doing centrifugation we can separate it from other impurities. And this is the only
purpose of the precipitation because immediately after the precipitation and washing we have to
dissolve it once again in the polar solution.
The precipitation process is just a modification in the DNA extraction protocol that facilitates
Ad
The alcohol guards the salt – DNA interaction, if it not shield, the precipitates will not form.
The higher dielectric constant water repels or hinders in the interaction of positive ions of the salt and
negative ions of the DNA so it is very difficult to dissolve DNA only using salt.
By adding the alcohol such as ethanol and isopropanol, the lower dielectric constant of the alcohol
protects the complex from water, and as we shake more the tube more positive charge and negative
charge of the DNA interacts with one another and we can get more precipitate.
Gsafe Green plus - Gsafe
Green plus
Ad
• Ammonium acetate = up to 5M
precipitation.
Ammonium acetate is one of the choices but not recommended for all types of DNA samples.
Sodium chloride and sodium acetate are the two best choices for DNA precipitation in which I
personally more favours the sodium acetate.
Ad
Ad
Alcohol used in the DNA precipitation:
Alcohol is a key factor in not only the DNA precipitation but also in DNA washing and DNA storing.
But the question arises in mind about methanol and other alcohol, why to use only ethanol and
isopropanol and not methanol?
Why methanol and other alcohol are not used in DNA precipitation?
Well, the DNA precipitation mechanism is solely dependent on the dielectric constant of the solvent.
Water 100 80
Methanol 68 33
Ethanol 78 ~24.3
Isopropanol 98 ~20.1
Take a close look at the methanol, the boiling point of methanol is 68°C and dielectric constant is 33.
The boiling point of methanol is lower than the ethanol and isopropanol while the dielectric constant is
higher.
Lower the dielectric point, higher the precipitation efficiency. We already discuss it why.
Furthermore, Boiling point of both the champions is higher that means it is safer to use.
In addition to this, the methanol is toxic to us, therefore, the Isopropanol and the ethanol are the two
best choices for the DNA precipitation.
However, if needed we can use methanol, it is totally fine and works nicely (not good for plasmid DNA
extraction).
with it?
Practically, the isopropanol also precipitates the salt too, means that impurities still remain in our DNA
and as we knew that the salts are the most dangerous inhibitors in the PCR reaction.
Hypothetical graph of dielectric constant vs the rate of DNA precipitation using different alcohol.
properly.
Let us start our protocol using two of the best chemicals for DNA precipitation, ethanol and sodium
acetate.
• Add the 1/10 volume of sodium acetate to the nucleic acid lysate solution.
• Add a doubled volume of pre-chilled ethanol.
• After the DNA precipitates appear (see the figure below) centrifuge the precipitated DNA at
• Now add 70% alcohol and wash the sample by inverting several times.
• Once the pellets dry properly, dissolve it in the TE buffer at pH nearly 8.2.
Gsafe Red plus - Gsafe Red plus
Safe&non-toxic, higher clarity&sensitivity than Gelred,
Glpbio is available from stock
Ad
The process of flocculation is very important for obtaining a higher yield of the precipitate. The
flocculation is achieved by chilling the sample.
At lower temperature, high “flok” or aggregates are obtained by addition of some of the precipitation
agents.
Addition of chilled ethanol facilitates higher flocculation in the process viz the higher precipitation of
DNA.
In some other protocols, if precipitates are not achieved, the sample is incubated at the lower
temperature (-20°C) or in the liquid nitrogen for some time to achieve precipitation.
Tips: For low DNA concentrate use 10 to 20 μg of glycogen and incubate the sample overnight at low
temperature increases the yield of DNA precipitation.
or
Ad
VISIT SITE
Add 10mM of MgCl2 to the precipitate and incubate for 1 to 1.5 hours at -20°C before centrifugation.
Both the modification greatly increases the yield of DNA precipitation of smaller fragment nucleic acid
or low concentrate nucleic acid.
For plasmid DNA or low concentrate DNA use this DNA precipitation protocol:
• Add the 1/10 volume of sodium acetate to the nucleic acid lysate solution.
• Now add 70% alcohol and wash the sample by inverting several times.
• Once the pellets dry properly, dissolve it in the TE buffer at pH nearly 8.2.
Tips: When you are using the isopropanol, do not freeze the sample or do not follow the chilling
Conclusion:
The world of DNA extraction is ruled by ready to use DNA extraction kits nowadays. However,
extracting DNA manually and visualizing the process of how DNA precipitate forms, is an amazing
experience for anyone.
Everyone should learn the manual methods of DNA extraction. Achieving pure DNA with good quantity
is an art. Learn manual things, it will enhance your knowledge and expertise.
Share this:
Share Article:
https://geneticeducation.co.in/a-quick-guide-on-dna-precipitation-and-dna-precipitation-protocol/
Tags: DNA precipitation DNA precipitation protocol Ethanol precipitation Salt in DNA precipitation
Dr Tushar Chauhan
15/04/2019
“Primer Dimer”: Zones DNA amplification by pairing with foe oligo
24/04/2019
PCR reaction: Ten secrets that nobody tells you
Leave a Reply
Enter your comment here...
Search in resources...
Visit Karyotypinghub.com
Our ebook
INSPIRON 5593
Legal Rollover
Buy Now
STUDENT OFFERS
INSPIRON 5593
Legal Rollover
Buy Now
Recent Posts
Categories
Books (1)
Chromosomes (2)
Comparisons (1)
COVID-19 (3)
Cytogenetics (10)
DNA (31)
Epigenetics (2)
Genes (2)
Genome (4)
Immunogenetics (3)
Mutation (6)
Replication (8)
Reviews (2)
RNA (7)
Transposons (9)
Trisomy (3)
Gsafe Red plus
Safe&non-toxic, higher clarity&sensitivity than Gelred, Glpbio is available from
stock
glpbio.com OPEN
report this ad
Home Archives
About Us
Select Month
Contact Us
Disclaimer
Privacy Policy
Enter your email address to subscribe to this blog and receive notifications of new posts by email.
Email Address
Subscribe
Genetic Education
© 2020 Genetic Education Inc. All rights reserved.
https://images.dmca.com/Badges/DMCABadgeHelper.min.js