Biotechnology Techniques

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Biotechnology Techniques

• Recombinant DNA
• DNA made by connecting DNA from two different
  sources.
• Example a human insulin
• gene inserted into bacterial
• DNA.
• Organisms containing
• recombinant DNA are called
• transgenic.

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Producing Recombinant DNA

• Utilizes restriction enzymes
• enzymes that cut specific sequences of DNA.
• The way that restriction enzymes cut DNA
  allows DNA from 2 different sources to be joined
  together.

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Sticky Ends

• Many restriction enzymes cut sequence in a way
  that produces single-stranded sticky ends.
• Any two DNA strands cut with the SAME
  restriction enzyme can bind together due to
  complementary pairing of sticky ends.

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Restriction Enzyme Video click once on image to
start
https//www.youtube.com/watch?vpiGOuud3f40
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Complementary sticky ends allow the DNA from 2
different sources to join together.
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Recombinant DNA video
https//www.youtube.com/watch?vTpmNfv1jKuA
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Pair Share

• Discuss with your table partner
• The definition of recombinant DNA
• How scientist are able to join two pieces of DNA
  from different sources together
• Some uses of recombinant DNA technology

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Vectors

• Vectors are a means by which foreign DNA can
  be transferred into a host cell.
• Common vectors
• Plasmids (small circular secondary chromosomes
  from bacteria)
• Viral DNA

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Bacterial Transformation

• Once a transgenic plasmid is produced, it can
  enter a bacterial cell under certain conditions.
  This uptake of foreign DNA by bacteria cells is
  called transformation.

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Plasmid Video Click once on image to start video
https//www.youtube.com/watch?vGNMJBMtKKWU
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Conditions for Transformation Treatment with
CaCl2 makes the bacterial cell wall more
permeable and help to allow the uptake of the
foreign DNA.
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Screening for Successive Transformation

• Not all the bacterial cells will successfully
  take in the foreign DNA with the desired gene.
• In order to be able to screen for successfully
  transformed cells, scientists often chose a
  plasmid vector with an antibiotic resistance gene
  and a host cell that is susceptible to the
  antibiotic.

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Transformed bacteria cell with antibiotic
resistance gene on plasmid
Bacteria cell that did NOT receive plasmid
Will be able to grow in the presence of the
antibiotic as well as on regular petri dishes.
Will only be able to grow on petri dishes that do
NOT contain the antibiotic.
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Pair Share

• Discuss with your table partner
• The definition of transformation (for bacteria)
• Screening of bacteria for successful
  transformation

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PCRPolymerase Chain Reaction

• Magnifies minute quantities of DNA so that
  they can be analyzed.
• Process
• DNA heated to separate strands (breaks weaker
  hydrogen bonds between nitrogen base pairs).
• Single strands incubated with DNA polymerase and
  free nucleotides to replicate.
• Process repeated multiple times.

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Pair Share

• Review with your table partner the process of
  polymerase chain reaction (PCR)
• What is the purpose of PCR
• What is the basic process
• How does the structure of DNA relate to how this
  process works (Hydrogen bonds, base pairs)