DNA Purification

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DNA Purification
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Definition

• DNA purification is a technique that removes
  impurities and unused reagents from samples after
  enzymatic reactions, such as PCR

www1.qiagen.com
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Quick Quiz

• The purpose(s) of DNA purification is to
• Remove excess reagents remaining from PCR
• Remove sample impurities
• Prepare the sample for downstream applications
  (i.e. sequencing)
• All of the above

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Non-examples

• DNA purification is not used to
• Isolate DNA from intact cells
• Change the concentration of DNA in a given volume
• Purify protein or RNA samples

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Downstream Applications

• Purified PCR products may be used in the
  following types of reactions
• Additional PCRs
• Sequencing
• Restriction digestion
• Ligation and
• transformation (Cloning)
• Labeling or hybridization

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Qiagen PCR Purification
Mix sample and buffer. Makes DNA in solution
ready to bind column.
Load onto column. Nucleic acids bind to
membrane. Other components flow through.
Wash Column. Removes any contaminants.
Elute DNA. DNA releases from column and is ready
for downstream applications.
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Quick Quiz

• In column purification, DNA is bound to the
  column membrane using a ______ salt solution.
• DNA is eluted off the membrane using a ______
  salt solution.
• High, low
• Low, high
• Iodized, non-iodized
• Saturated, supersaturated

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Quick Quiz

• Which list of activities is in the correct order?
  
• DNA extraction, DNA purification, PCR, DNA
  sequencing
• DNA purification, DNA extraction, PCR, DNA
  sequencing
• DNA extraction, PCR, DNA purification, DNA
  sequencing
• DNA extraction, PCR, DNA sequencing, DNA
  purification

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• END

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Resources

• Modified QIAquick protocol PCR Purification
  handout
• Qiagen QIAquick PCR purification protocol

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California State Chemistry Standards

• Grade 8
• 6c. Living organisms have many different kinds
  of molecules
• Grades 9-12
• 6. Solutions are homogenous mixtures of two or
  more substances
• 7. Energy is exchanged or transformed in all
  chemical reactions and physical changes of matter
• 8. Chemical reaction rates depend on factors that
  influence the frequency of collision of reactant
  molecules

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California State Biology Standards

• Grade 7
• 2e. DNA is the genetic material of living
  organisms and is located in the chromosomes of
  each cell
• 3a. Genetic variation and environmental factors
  are causes of evolution and diversity of
  organisms
• Grades 9-12
• 2. Mutation and sexual reproduction lead to
  genetic variation in a population
• 7. The frequency of an allele in a gene pool of
  a population depends on many factors and may be
  stable or unstable over time

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California State Investigation and
ExperimentationStandards

• Grades 7, 9-12
• a/b. Select and use appropriate tools and
  technology to perform tests, collect data,
  analyze relationships, and display data
• Grade 8
• a. Plan and conduct a scientific investigation
  to test a hypothesis

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National Standards

• Grades 6-12
• Content Standard A Science as Inquiry
•  
• Content Standard C Life Science
• Content Standard E Science and Technology