Biotechnology and Genetic Engineering

1. Watch this video

2. Study this summary

Recombinant DNA

  • Definition: DNA combined from more than one source (e.g., bacterial DNA with human DNA).
  • Creation:
    • Use restriction enzymes to cut DNA at specific sequences (restriction sites), creating fragments with sticky ends.
    • Sticky ends form hydrogen bonds with complementary sequences.
    • Use DNA ligase to create sugar-phosphate bonds, joining fragments into recombinant DNA.

Recombinant Plasmids

  • Process:
    1. Extract a plasmid from bacteria and cut it with restriction enzymes.
    2. Use the same restriction enzymes to cut a human gene (introns removed).
    3. Combine the plasmid and human gene; sticky ends bind via hydrogen bonds.
    4. Use DNA ligase to seal the bonds, forming a recombinant plasmid.
    5. Insert the plasmid into bacteria via transformation.
  • Example: Bacteria engineered to produce human insulin.

Solving the Problem of Introns When Making Recombinant DNA

  • Problem: Bacteria cannot process introns, leading to nonfunctional proteins.
  • Solution:
    1. Reverse-engineering: Use the genetic code to determine DNA sequences for desired proteins.
    2. Using mRNA: Extract mRNA (introns already removed), then use reverse transcriptase to create complementary DNA (cDNA).

Gel Electrophoresis

  • Definition: Technique to sort DNA fragments by size and charge.
  • Process:
    • DNA is placed in a gel within an electric field.
    • Negatively charged DNA moves towards the positive electrode; smaller fragments travel farther.
    • Use: Analyze DNA fragment patterns for forensics, genetic studies, or restriction mapping.

Restriction Mapping

  • Definition: Mapping the locations of restriction sites on a DNA molecule.
  • Example: Cutting a plasmid with restriction enzymes to produce fragments of specific sizes, which are then analyzed via gel electrophoresis.

Polymerase Chain Reaction (PCR)

  • Definition: A cell-free method for cloning DNA in a test tube.
  • Requirements:
    • DNA sample, primers, heat-resistant DNA polymerase, and free nucleotides.
  • Steps:
    1. Heat DNA to separate strands.
    2. Cool to allow primers to bind.
    3. DNA polymerase synthesizes new DNA.
  • Outcome: Each cycle doubles DNA quantity; 30 cycles yield a billionfold amplification.
  • Applications: Forensics, research, and diagnostics.

DNA Sequencing

  • Definition: Determining the precise order of nucleotides (A, T, C, G) in DNA.
  • Uses:
    • Identify proteins an organism can produce.
    • Infer evolutionary relationships.
    • Analyze cancer mutations.
    • Study and monitor viral variants (e.g., SARS-CoV-2).
    • Forensics: Identify suspects, exonerate individuals, and resolve paternity disputes.

3. Master these flashcards

[qdeck]
[h]Biotechnology (AP Biology Topic 6.8)

[q]What is recombinant DNA?
[a]DNA combined from more than one source, such as bacterial DNA with human DNA.

[q]How is recombinant DNA created?
[a]1. Use restriction enzymes to cut DNA at specific sequences, creating fragments with sticky ends.
2. Sticky ends form hydrogen bonds with complementary sequences.
3. Use DNA ligase to join fragments into recombinant DNA.

[q]What are the steps to create a recombinant plasmid?
[a]1. Extract a plasmid from bacteria and cut it with restriction enzymes.
2. Cut a human gene (introns removed) with the same restriction enzymes.
3. Combine plasmid and human gene; sticky ends bind via hydrogen bonds.
4. Use DNA ligase to seal the bonds, forming a recombinant plasmid.
5. Insert the plasmid into bacteria via transformation.

[q]Give an example of recombinant DNA used in medicine.
[a]Bacteria engineered to produce human insulin.

[q]Why must introns be removed before transferring a human gene to bacteria?
[a]Bacteria cannot process introns, so leaving them in would lead to a nonfunctional protein.

[q]What are two methods to remove introns for recombinant DNA?
[a]1. Reverse-engineering the DNA sequence using the protein’s amino acid sequence.
2. Extracting mRNA and using reverse transcriptase to create complementary DNA (cDNA).

[q]What is gel electrophoresis?
[a]A technique used to sort DNA fragments by size and charge.

[q]How does gel electrophoresis work?
[a]1. DNA is placed in a gel within an electric field.
2. Negatively charged DNA moves towards the positive electrode.
3. Smaller fragments travel farther through the gel.
4. Fragments are sorted by size.

[q]What is restriction mapping?
[a]Mapping the locations of restriction sites on a DNA molecule by analyzing fragment patterns using gel electrophoresis.

[q]What is PCR (polymerase chain reaction)?
[a]A cell-free method for cloning DNA in a test tube.

[q]What are the requirements for PCR?
[a]1. DNA sample
2. Primers
3. Heat-resistant DNA polymerase
4. Free nucleotides

[q]What are the steps of PCR?
[a]1. Heat DNA to separate strands.
2. Cool to allow primers to bind.
3. DNA polymerase synthesizes new DNA.
4. Each cycle doubles the DNA quantity.

[q]What are some applications of PCR?
[a]Forensics, research, diagnostics, and amplifying DNA from small samples.

[q]What is DNA sequencing?
[a]Determining the precise order of nucleotides (A, T, C, G) in DNA.

[q]What are some uses of DNA sequencing?
[a]1. Identifying proteins an organism can produce.
2. Inferring evolutionary relationships.
3. Analyzing cancer mutations.
4. Monitoring viral variants (e.g., SARS-CoV-2).
5. Forensics, including identifying suspects and resolving paternity disputes.

[/qdeck]

4. Tackle these quizzes

4.1. Genetic Engineering Overview

[qwiz random = “true”]

[h]Genetic Engineering Overview

[i]


[q]Genetically engineered insulin is used to treat [hangman].

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[q] In the diagram below, which letter represents the main bacterial chromosome?

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[c]IEVudGVyIHdvcmQ=[Qq]

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[q] In the diagram below, which letter represents an unmodified plasmid?

[textentry single_char=”true”]

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[f]IE5pY2Ugam9iLsKgICYjODIyMDtDJiM4MjIxOyByZXByZXNlbnRzIGFuIHVubW9kaWZpZWQgcGxhc21pZC4=[Qq]

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[q] In the diagram below, which letter represents the human insulin gene?

[textentry single_char=”true”]

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[q] In the diagram below, a plasmid with recombinant DNA is shown at

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[q] In the diagram below, which letter indicates the reproduction of genetically engineered offspring?

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[q] In the diagram below, which letter indicates the production of genetically engineered insulin?

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[q]DNA that is a combination of DNA from two different sources is called [hangman] DNA.

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[q]In the diagram below, letter “b” represents [hangman]. Number 3 shows glucose entering into the cell by a type of membrane transport called [hangman] [hangman].

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[q]Insulin is a [hangman] hormone, the function of which is to reduce the amount of [hangman] in the bloodstream.

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[/qwiz]

4.2. Genetic Engineering Quiz 2

The following quiz will help you master everything you’ve read above.

[qwiz random = “true”] [h]

Genetic Engineering Quiz 2

[i]

[q] In the diagram below, a eukaryotic gene of interest is indicated by
[textentry single_char=”true”]

 

[c]ID E=[Qq]

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[q] In the diagram below, restriction enzymes are at which number?

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[q] In the diagram below, a non-recombinant plasmid is indicated by
[textentry single_char=”true”]

[c]ID M=[Qq]

[f]V2F5IHRvIGdvLiBUaGUgdW5tb2RpZmllZCBwbGFzbWlkIGlzIGluZGljYXRlZCBieSAmIzgyMjA7My4mIzgyMjE7[Qq]

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[q] In the diagram below, DNA ligase is indicated by

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[c]ID Q=[Qq]

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[q] In the diagram below, fragments of DNA with “sticky ends” are represented by
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[f]IEF3ZXNvbWUhIFRoZSBzdGlja3kgZW5kcyBhcmUgcmVwcmVzZW50ZWQgYnkgJiM4MjIwOzUuJiM4MjIxOw==[Qq]

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[q] In the diagram below, a recombinant plasmid is shown at
[textentry single_char=”true”]

[c]ID Y=[Qq]

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[q] Organisms that have been modified to contain DNA from more than one species are called[hangman]. Often, these organisms are referred to as genetically [hangman] organisms, or GMOs.

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[q] Enzymes used to cut DNA are called[hangman]  enzymes.

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[q] DNA [hangman]is the enzyme that connects DNA fragments by creating sugar-phosphate bonds.

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[q]  [hangman]  DNA is DNA that has been pieced together from multiple sources.

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[q] In the diagram below, a restriction site is shown at letter

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[q] In the diagram below, a restriction enzyme is shown at

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[q] In the diagram below, “sticky ends” are shown at

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[q] In the diagram below, gaps that need to be sealed together by DNA ligase are shown at

[textentry single_char=”true”]

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[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLCB0aGF0JiM4MjE3O3Mgbm90IGNvcnJlY3Qu[Qq]

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[q] In the diagram below, DNA ligase is shown at

[textentry single_char=”true”]

[c]IE k=[Qq]

[f]IENvcnJlY3QhICYjODIyMDtJJiM4MjIxOyByZXByZXNlbnRzIGxpZ2FzZS7CoA==[Qq]

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[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50LiBMaWdhc2Ugc2VhbHMgdGhlIGdhcHMgdGhhdCBhcmUgc2hvd24gYXQgJiM4MjIwO0guJiM4MjIxOw==[Qq]

[q] In the diagram below, completed recombinant DNA is shown at

[textentry single_char=”true”]

[c]IE o=[Qq]

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[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLCB0aGF0JiM4MjE3O3Mgbm90IGNvcnJlY3Qu[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50LiBSZWNvbWJpbmFudCBETkEgY29tYmluZXMgdGhlIGZyYWdtZW50cyBhdCAmIzgyMjA7RCYjODIyMTsgYW5kICYjODIyMDtGLiYjODIyMTs=[Qq]

[q] DNA made from RNA using reverse transcriptase is known as [hangman] DNA.

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[q multiple_choice=”true”] The enzyme responsible for creating sugar-phosphate bonds between DNA fragments is/are

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[c]IFJldmVyc2UgdHJhbnNjcmlwdGFzZQ==[Qq]

[f]IE5vLiBSZXZlcnNlIHRyYW5zY3JpcHRhc2UgdGFrZXMgUk5BIGFuZCByZXZlcnNlIHRyYW5zY3JpYmVzIGl0IGludG8gRE5BLiBIZXJlJiM4MjE3O3MgYSBoaW50OiBjcmVhdGluZyB0aGVzZSBzdWdhci1waG9zcGhhdGUgYm9uZHMgaXMgY2FsbGVkICYjODIyMDtsaWdhdGlvbi4mIzgyMjE7[Qq]

[c]IFJlc3RyaWN0aW9uIGVuenltZXM=[Qq]

[f]IE5vLiBSZXN0cmljdGlvbiBlbnp5bWVzIGFyZSByZXNwb25zaWJsZSBmb3IgY3V0dGluZyBETkEgaW50byBmcmFnbWVudHMuIEhlcmUmIzgyMTc7cyBhIGhpbnQ6IGNyZWF0aW5nIHRoZXNlIHN1Z2FyLXBob3NwaGF0ZSBib25kcyBpcyBjYWxsZWQgJiM4MjIwO2xpZ2F0aW9uLiYjODIyMTs=[Qq]

[c]IEROQSBs aWdhc2U=[Qq]

[f]IENvcnJlY3QhIEROQSBMaWdhc2UgY3JlYXRlcyBzdWdhci1waG9zcGhhdGUgYm9uZHMgYmV0d2VlbiBhZGphY2VudCBmcmFnbWVudHMgb2YgRE5BLg==[Qq]

[q multiple_choice=”true”] The enzyme that can synthesize DNA from an RNA template is

[c]IEROQSBwb2x5bWVyYXNl[Qq]

[f]IE5vLiBETkEgcG9seW1lcmFzZSBpcyBhIGtleSBlbnp5bWUgcmVzcG9uc2libGUgZm9yIHN5bnRoZXNpemluZyBuZXcgRE5BLg==[Qq]

[c]IFJldmVyc2UgdHJh bnNjcmlwdGFzZQ==[Qq]

[f]IE5pY2Ugam9iLiBSZXZlcnNlIHRyYW5zY3JpcHRhc2UgdGFrZXMgUk5BIGFuZCByZXZlcnNlIHRyYW5zY3JpYmVzIGl0IGludG8gRE5BLg==[Qq]

[c]IFJlc3RyaWN0aW9uIGVuenltZXM=[Qq]

[f]IE5vLiBSZXN0cmljdGlvbiBlbnp5bWVzIGFyZSByZXNwb25zaWJsZSBmb3IgY3V0dGluZyBETkEgaW50byBmcmFnbWVudHM=[Qq]

[c]IEROQSBsaWdhc2U=[Qq]

[f]IE5vLiBETkEgTGlnYXNlIGNyZWF0ZXMgc3VnYXItcGhvc3BoYXRlIGJvbmRzIGJldHdlZW4gYWRqYWNlbnQgZnJhZ21lbnRzIG9mIEROQS4=[Qq]

[q multiple_choice=”true”] The enzyme that can cut DNA into fragments is

[c]IEROQSBwb2x5bWVyYXNl[Qq]

[f]IE5vLiBETkEgcG9seW1lcmFzZSBpcyBhIGtleSBlbnp5bWUgcmVzcG9uc2libGUgZm9yIHN5bnRoZXNpemluZyBuZXcgRE5BLg==[Qq]

[c]IFJldmVyc2UgdHJhbnNjcmlwdGFzZQ==[Qq]

[f]IE5vLiBSZXZlcnNlIHRyYW5zY3JpcHRhc2UgdGFrZXMgUk5BIGFuZCByZXZlcnNlIHRyYW5zY3JpYmVzIGl0IGludG8gRE5BLg==[Qq]

[c]IFJlc3RyaWN0aW 9uIGVuenltZXM=[Qq]

[f]IEV4Y2VsbGVudC4gUmVzdHJpY3Rpb24gZW56eW1lcyBhcmUgcmVzcG9uc2libGUgZm9yIGN1dHRpbmcgRE5BIGludG8gZnJhZ21lbnRz[Qq]

[c]IEROQSBsaWdhc2U=[Qq]

[f]IE5vLiBETkEgTGlnYXNlIGNyZWF0ZXMgc3VnYXItcGhvc3BoYXRlIGJvbmRzIGJldHdlZW4gYWRqYWNlbnQgZnJhZ21lbnRzIG9mIEROQS4=[Qq]

[q] In the diagram below, a restriction enzyme is represented by
[textentry single_char=”true”]

[c]IG U=[Qq]

[f]IENvcnJlY3QhIExldHRlciAmIzgyMjA7ZSYjODIyMTsgaXMgYSByZXN0cmljdGlvbiBlbnp5bWUu[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLg==[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50LiBSZXN0cmljdGlvbiBlbnp5bWVzIGN1dCBvcGVuIEROQS4=[Qq]

[q] In the diagram below, HindIII must be a(n) [hangman] enzyme.

[c]IHJlc3RyaWN0aW9u[Qq]

[f]IEV4Y2VsbGVudCE=[Qq]

[q] In the diagram below, the DNA bases with green arrows represent[hangman] ends.

[c]IHN0aWNreQ==[Qq]

[f]IEV4Y2VsbGVudCE=[Qq]

[q] In the diagram below, a plasmid that’s been treated with a restriction enzyme is shown at

[textentry single_char=”true”]

[c]IG U=[Qq]

[c]WWVzISBMZXR0ZXIgJiM4MjIwO2UmIzgyMjE7IHNob3dzIGEgcGxhc21pZCB0aGF0JiM4MjE3O3MgYmVlbiBjdXQgb3BlbiBieSBhIHJlc3RyaWN0aW9uIGVuenltZS4=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50LiBBIHBsYXNtaWQgdGhhdCA=aGFzbiYjODIxNzt0IGJlZW4gdHJlYXRlZCB3aXRoIGEgcmVzdHJpY3Rpb24gZW56eW1lIGlzIHNob3duIGF0ICYjODIyMDthLiYjODIyMTs=[Qq]

[q] In the diagram below, recombinant DNA first appears at

[textentry single_char=”true”]

[c]IG Y=[Qq]

[f]IFllcyEgJiM4MjIwO0YmIzgyMjE7IGlzIGEgcmVjb21iaW5hbnQgcGxhc21pZC4=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50LiBMb29rIGZvciB0aGUgZmlyc3Qgb2NjdXJyZW5jZSBvZiBodW1hbiBETkEgY29tYmluZWQgd2l0aCBwbGFzbWlkIEROQS4=[Qq]

[q] In the diagram below, the first occurrence of a transgenic organism is at

[textentry single_char=”true”]

[c]IG c=[Qq]

[f]WWVzISBMZXR0ZXIgJiM4MjIwO2cmIzgyMjE7IHNob3dzIGEgcmVjb21iaW5hbnQgb3JnYW5pc20u[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50LiBMb29rIGZvciBhIGNlbGwgdGhhdCBoYXMgYm90aCBiYWN0ZXJpYWwgRE5BIGFuZCBodW1hbiBETkEu[Qq]

[x]

[restart]

[/qwiz]

 

4.3. Interpreting Gel Electrophoresis Results

[qwiz]

[h]Results of electrophoresis

[q] What’s the smallest fragment in the gel shown below?

[textentry single_char=”true”]

[c]ID Y=[Qq]

[f]TmljZS4gJiM4MjIwOzYmIzgyMjE7IHdvdWxkIGhhdmUgdG8gYmUgdGhlIHNtYWxsZXN0IGZyYWdtZW50Lg==[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBJbiBnZWwgZWxlY3Ryb3Bob3Jlc2lzLCB0aGUgc21hbGxlc3QgZnJhZ21lbnQgaXMgZ29pbmcgdG8gbW92ZSB0aGUgZmFzdGVzdCwgYW5kIGluIGEgZ2l2ZW4gcGVyaW9kLCB0aGUgZmFydGhlc3Qu[Qq]

[q] What’s the largest fragment in the gel shown below?

[textentry single_char=”true”]

[c]ID E=[Qq]

[f]RXhjZWxsZW50OiBOdW1iZXIgMSwgd2hpY2ggbW92ZWQgdGhlIHNob3J0ZXN0IGRpc3RhbmNlLCB3b3VsZCBoYXZlIHRoZSBiZSB0aGUgbGFyZ2VzdCBmcmFnbWVudC4=[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBJbiBnZWwgZWxlY3Ryb3Bob3Jlc2lzLCB0aGUgbGFyZ2VzdCBmcmFnbWVudCBpcyBnb2luZyB0byBtb3ZlIHRoZSBzbG93ZXN0LCBhbmQgaW4gYSBnaXZlbiBwZXJpb2QsIHRoZSBzbWFsbGVzdCBkaXN0YW5jZS4=[Qq]

[q] Assuming that all three samples below consist of linear DNA (not a circular plasmid) and were exposed to the same restriction enzyme, which sample had one restriction site?

[textentry single_char=”true”]

[c]IE I=[Qq]

[f]TmljZTogU2FtcGxlIEIgd2FzIGN1dCBpbnRvIHR3byBmcmFnbWVudHMuIFRoYXQgbWVhbnMgdGhhdCB0aGUgRE5BIGluIHNhbXBsZSBCIGhhZCBvbmUgcmVzdHJpY3Rpb24gc2l0ZSwgYW5kIGEgcmVzdHJpY3Rpb24gZW56eW1lIG1hZGUgYSBjdXQgYXQgdGhhdCBzaXRlLCBkaXZpZGluZyB0aGUgRE5BIGludG8gdHdvIGZyYWdtZW50cy4=[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50LiBJZiB0aGUgRE5BIGhhcyBvbmUgcmVzdHJpY3Rpb24gc2l0ZSwgaXQmIzgyMTc7bGwgYmUgY3V0IGludG8gdHdvIGZyYWdtZW50cy4=[Qq]

[q] Assuming that all three samples below consist of linear DNA (not a circular plasmid) which sample had two restriction sites?

[textentry single_char=”true”]

[c]IE M=[Qq]

[f]R3JlYXQgam9iOiBTYW1wbGUgQyB3YXMgY3V0IGludG8gdGhyZWUgZnJhZ21lbnRzLiBUaGF0IG1lYW5zIHRoYXQgdGhlIEROQSBpbiBzYW1wbGUgQyBoYWQgdHdvIHJlc3RyaWN0aW9uIHNpdGVzLiBUaGUgcmVzdHJpY3Rpb24gZW56eW1lIG1hZGUgYSBjdXQgYXQgZWFjaCBvZiB0aG9zZSBzaXRlcywgZGl2aWRpbmcgdGhlIEROQSBpbnRvIHRocmVlIGZyYWdtZW50cy4=[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50LiBJZiB0aGUgRE5BIGhhcyBvbmUgcmVzdHJpY3Rpb24gc2l0ZSwgaXQmIzgyMTc7bGwgYmUgY3V0IGludG8gdHdvIGZyYWdtZW50cy4gSWYgaXQgaGFzIHR3byByZXN0cmljdGlvbiBzaXRlcywgaXQmIzgyMTc7bGwgYmUgY3V0IGludG8gdGhyZWUgZnJhZ21lbnRzLg==[Qq]

[q] Assuming that all three samples below consist of linear DNA (not a circular plasmid) which sample had no restriction sites for that restriction enzyme?

[textentry single_char=”true”]

[c]IE E=[Qq]

[f]VGVycmlmaWMuIFNhbXBsZSBBIGlzIG9uZSBmcmFnbWVudCwgcmVwcmVzZW50ZWQgYnkgdGhlIGxpbmUgYXQgMS4gVGhhdCBtZWFucyB0aGF0IHRoZSBETkEgaW4gc2FtcGxlIEEgaGFkIG5vIHJlc3RyaWN0aW9uIHNpdGVzLCBhbmQgd2FzbiYjODIxNzt0IGN1dCBhcGFydC4=[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50LiBJZiB0aGUgRE5BIGhhcyBhbnkgcmVzdHJpY3Rpb24gc2l0ZXMsIGl0JiM4MjE3O2xsIGJlIGN1dCBpbnRvIHNtYWxsZXIgZnJhZ21lbnRzLiBJZiBpdCByZW1haW5zIG9uZSBwaWVjZSB0aGF0IG1lYW5zIHRoYXQgbm90aGluZyBjdXQgaXQgYXBhcnQu[Qq]

[/qwiz]

 

4.4. Plasmid Mapping

[qwiz]

[h]Restriction Sites and Restriction Fragments in Plasmid DNA

[i]

Plasmids are shown at number 2

[q] If the plasmid below is treated with EcoRI and HindIII, the result will be[hangman] restriction fragment(s). The shorter one will be [hangman] bp long, and the longer one will be  [hangman] bp long.

[c]IDI=[Qq]

[f]IEdvb2Qh[Qq]

[c]IDUwMA==[Qq]

[f]IEdyZWF0IQ==[Qq]

[c]IDM1MDA=[Qq]

[f]IEV4Y2VsbGVudCE=[Qq]

[q] If the plasmid below is treated with BamH I and HindIII, the result will be [hangman] restriction fragment(s). The shorter one will be [hangman] bp long, and the longer one will be  [hangman] bp long.

[c]IDI=[Qq]

[f]IEdyZWF0IQ==[Qq]

[c]IDE1MDA=[Qq]

[f]IENvcnJlY3Qh[Qq]

[c]IDI1MDA=[Qq]

[f]IEV4Y2VsbGVudCE=[Qq]

[q multiple_choice = “true] The image to the left, below, shows a 20 kb plasmid with several restriction sites. The image on the right shows the results of electrophoresis following various combinations of restriction enzymes. Which lane shows the gel that would result if the plasmid were digested with BamH I?

[c]IEHCoCDCoCDCoCA=[Qq][c]IELCoCDC oCDCoMKg[Qq][c]IEM=

Cg==[Qq]

[f]IE5vLiBUaGUgc2luZ2xlIGxpbmUgaW4gbGFuZSAmIzgyMjA7QSYjODIyMTsgaW5kaWNhdGVzIHRoYXQgb25seSBvbmUgZnJhZ21lbnQgd291bGQgcmVzdWx0IGFuZCB0aGF0IHRoaXMgZnJhZ21lbnQgd291bGQgYmUgMjAga2IgaW4gc2l6ZS4gSWYgeW91IGN1dCB0aGlzIHBsYXNtaWQgd2l0aCBCYW1IMSwgeW91JiM4MjE3O2xsIG1ha2UgMyBjdXRzLiBOb3csIGp1c3QgZmlndXJlIG91dCBob3cgYmlnIGVhY2ggb2YgdGhlIGZyYWdtZW50cyB3b3VsZCBiZSwgYW5kIHlvdSYjODIxNztsbCBoYXZlIHlvdXIgYW5zd2VyLg==[Qq]

[f]IEV4Y2VsbGVudC4gQ3V0dGluZyB0aGlzIHBsYXNtaWQgd2l0aCBCYW1ISSB3aWxsIHJlc3VsdCBpbiB0aHJlZSBmcmFnbWVudHMsIGFuZCB0aGV5JiM4MjE3O2xsIGJlIDExIGtiLCA2IGtiLCBhbmQgMyBLYiBpbiBzaXplLCB3aGljaCBpcyBqdXN0IHdoYXQgaXMgc2hvd24gaW4gdGhpcyBnZWwu[Qq]

[f]IE5vLiBUaGVyZSBhcmUgdGhyZWUgbGluZXMgaW4gbGFuZSBDLCBhbmQgb25lIG9mIHRob3NlIGNvbnNpc3RzIG9mIHR3byBmcmFnbWVudHMuIFRoZSBmaXJzdCBsaW5lIGNvbnNpc3RzIG9mIGEgcmVzdHJpY3Rpb24gZnJhZ21lbnQgdGhhdCYjODIxNztzIDgga2IgbG9uZzsgdGhlIHNlY29uZCBoYXMgYSBmcmFnbWVudCB0aGF0JiM4MjE3O3MgNiBrYiBsb25nLCBhbmQgdGhlIHRoaXJkIGNvbnNpc3RzIG9mIHR3byBmcmFnbWVudHMsIGJvdGggb2Ygd2hpY2ggYXJlIDMga2IuIFdoaWNoIGVuenltZXMgd291bGQgcHJvZHVjZSB0aGF0IHBhdHRlcm4gb2YgZnJhZ21lbnRzPw==[Qq]

[q multiple_choice = “true] The image to the left, below, shows a 20 kb plasmid with several restriction sites. The image on the right shows the results of electrophoresis following various combinations of restriction enzymes. Which lane shows the gel that would result if the plasmid were digested with EcoRI?

[c]IEHCoCDC oCDCoA==[Qq][c]IELCoCDCoCDCoA==[Qq][c]IEM=

Cg==[Qq]

[f]IE5pY2UgSm9iISBUaGUgc2luZ2xlIGxpbmUgaW4gbGFuZSAmIzgyMjA7QSYjODIyMTsgaW5kaWNhdGVzIHRoYXQgb25seSBvbmUgZnJhZ21lbnQgd291bGQgcmVzdWx0IGFuZCB0aGF0IHRoaXMgZnJhZ21lbnQgd291bGQgYmUgMjAga2IgaW4gc2l6ZS4=[Qq]

[f]IE5vLiBUaGVyZSYjODIxNztzIG9ubHkgb25lIHJlc3RyaWN0aW9uIHNpdGUgcmVjb2duaXplZCBieQ==IEVjbw==IFJJLiBJZiB0aGUgcGxhc21pZCBpcyBjdXQgb3ZlciBhdCB0aGF0IHBvaW50LCBob3cgbWFueSBmcmFnbWVudHMgd291bGQgcmVzdWx0Pw==[Qq]

[f]IE5vLiBUaGVyZSBhcmUgdGhyZWUgbGluZXMgaW4gbGFuZSBDLCBhbmQgb25lIG9mIHRob3NlIGNvbnNpc3RzIG9mIHR3byBmcmFnbWVudHMuIEJ1dCB0aGVyZSYjODIxNztzIG9ubHkgb25lIHJlc3RyaWN0aW9uIHNpdGUgcmVjb2duaXplZCBieQ==IEVjbw==IFJJLiBJZiB0aGUgcGxhc21pZCBpcyBjdXQgb3ZlciBhdCB0aGF0IHBvaW50LCBob3cgbWFueSBmcmFnbWVudHMgd291bGQgcmVzdWx0Pw==[Qq]

[q multiple_choice = “true] The image to the left, below, shows a 20 kb plasmid with several restriction sites. The image on the right shows the results of electrophoresis following various combinations of restriction enzymes. Which lane shows the gel that would result if the plasmid were digested with Bam HI and EcoRI?

[c]IEHCoCDCoCDCoA==[Qq][c]IELCoCDCoCDCoA==[Qq][c]IE M=

Cg==[Qq]

[f]IE5vLiBUaGUgc2luZ2xlIGxpbmUgaW4gbGFuZSAmIzgyMjA7QSYjODIyMTsgaW5kaWNhdGVzIHRoYXQgdGhlcmUmIzgyMTc7cyBvbmx5IG9uZSBmcmFnbWVudCBhbmQgdGhhdCB0aGlzIGZyYWdtZW50IHdvdWxkIGJlIDIwIGtiIGluIHNpemUuIEhvdyBjb3VsZCB0aGF0IG9jY3VyIGlmIGJvdGggQmFtIEhJIA==YW5kIA==[Qq]Eco R I were used to digest the plasmid?

[f]IE5vLiBMYW5lIEIgaW5kaWNhdGVzIG9uZSBmcmFnbWVudCBvZiAxMSBrYiwgYSBzZWNvbmQgb2YgNiBrYiwgYW5kIGEgM3JkIG9mIDMga2Iu[Qq]

[f]IEV4Y2VsbGVudC4gVGhlcmUgYXJlIHRocmVlIGxpbmVzIGluIGxhbmUgQywgYW5kIG9uZSBvZiB0aG9zZSBjb25zaXN0cyBvZiB0d28gZnJhZ21lbnRzLiBUaGF0JiM4MjE3O3MgZXhhY3RseSB3aGF0IHlvdSYjODIxNztkIGV4cGVjdCBpZiBib3RowqA=QmFtIEhJIA==YW5kIA==[Qq]Eco R I were used to digest the plasmid.

[/qwiz]

4. PCR: Checking Understanding

Complete the following quiz.

[qwiz random = “true” style=”width: 650px !important;”]

[h]PCR Quiz

[i]

[q] In the diagram below, which number or letter indicates the target DNA that’s about to be amplified?

 

[textentry single_char=”true”]

[c]IG E=[Qq]

[f]IEV4Y2VsbGVudCEgTGV0dGVyICYjODIyMDthJiM4MjIxOyBpcyB0aGUgdGFyZ2V0IEROQS4=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBKdXN0IGlkZW50aWZ5IHdoYXQmIzgyMTc7cyBiZWluZyBjb3BpZWQsIGFuZCB5b3UmIzgyMTc7bGwgaGF2ZSB5b3VyIGFuc3dlci4=[Qq]

[c]IEVudGVyIGxldHRlcg==[Qq]

[f]IE5vLCB0aGF0JiM4MjE3O3Mgbm90IGNvcnJlY3Qu[Qq]

[q] In the diagram below, which letter indicates free nucleotides?

[textentry single_char=”true”]

[c]IG I=[Qq]

[f]IEV4Y2VsbGVudCEgTGV0dGVyICYjODIyMDtiJiM4MjIxOyBzaG93cyBmcmVlIG51Y2xlb3RpZGVzLsKg[Qq]

[c]ICo=[Qq]

[f]IE5vLiBMb29rIGZvciB0aGUgbW9ub21lcnMsIG9yIHVuaXRzLCB0aGF0IGFyZSBnb2luZyB0byBiZSB1c2VkIHRvIGJ1aWxkIG5ldyBETkEu[Qq]

[q] In the diagram below, which number or letter indicates the primer before it binds with the template DNA?

[textentry single_char=”true”]

[c]IG M=[Qq]

[f]IENvcnJlY3QhIExldHRlciAmIzgyMjA7YyYjODIyMTsgcmVwcmVzZW50cyBhbiB1bmJvdW5kIHByaW1lci4=[Qq]

[c]ICo=[Qq]

[f]IE5vLiBMb29rIGZvciBhIHNob3J0IHN0cmluZyBvZiBudWNsZW90aWRlcyB0aGF0IHdpbGwgYmluZCBhdCB0aGUgc3RhcnQgb2YgdGhlIHRhcmdldCBETkEu[Qq]

[q] In the diagram below, which letter indicates the DNA after it’s been denatured?

[textentry single_char=”true”]

[c]IG Q=[Qq]

[f]IENvcnJlY3QhIExldHRlciAmIzgyMjA7ZCYjODIyMTsgcmVwcmVzZW50cyBkZW5hdHVyZWQgRE5BIChub3cgdHdvIA==c2luZ2xlIHN0cmFuZHMpLsKg[Qq]

[c]ICo=[Qq]

[f]IE5vLiBEZW5hdHVyYXRpb24gYnJlYWtzIHRoZSBoeWRyb2dlbiBib25kcyB0aGF0IGhvbGQgdGhlIGRvdWJsZSBoZWxpeCB0b2dldGhlci4gT25jZSB0aG9zZSBib25kcyBhcmUgYnJva2VuLCB3aGF0IHdpbGwgdGhlIEROQSBsb29rIGxpa2U/[Qq]

[q] In the diagram below, which letter shows a primer binding with the template DNA?

[textentry single_char=”true”]

[c]IG U=[Qq]

[f]IENvcnJlY3QhIExldHRlciAmIzgyMjA7ZSYjODIyMTsgc2hvd3MgYSBwcmltZXIgYmluZGluZyB3aXRoIHRoZSB0ZW1wbGF0ZSBETkEu[Qq]

[c]ICo=[Qq]

[f]IE5vLiBMb29rIGZvciBhIHNtYWxsIHNlcXVlbmNlIG9mIG51Y2xlb3RpZGVzIHRoYXQmIzgyMTc7cyBjb21wbGVtZW50YXJ5IHdpdGggbnVjbGVvdGlkZXMgYXQgdGhlIDMmIzgyNDI7IGVuZCBvZiB0aGUgdGVtcGxhdGUgc3RyYW5kLg==[Qq]

[q] In the diagram below, which letter indicates newly synthesized DNA?

[textentry single_char=”true”]

[c]IG Y=[Qq]

[f]IENvcnJlY3QhICYjODIyMDtGJiM4MjIxOyBzaG93cyBuZXdseSBzeW50aGVzaXplZCBETkEu[Qq]

[c]ICo=[Qq]

[f]IE5vLiBMb29rIGZvciB0aGUgYXNzZW1ibHkgb2YgdGhlIGluZGl2aWR1YWwgbnVjbGVvdGlkZXMgKHNob3duIGF0ICYjODIyMDtiJiM4MjIxOykgaW50byBhIGNvbm5lY3RlZCBzdHJhbmQu[Qq]

[q] In the diagram below, which number or letter indicates DNA polymerase?

[textentry single_char=”true”]

[c]IE c=[Qq]

[f]IENvcnJlY3QhICYjODIyMDtHJiM4MjIxOyBzaG93cyBETkEgcG9seW1lcmFzZQ==[Qq]

[c]ICo=[Qq]

[f]IE5vLiBETkEgcG9seW1lcmFzZSBpcyB0aGUgZW56eW1lIHRoYXQgbWFrZXMgdGhlIG5ldyBETkEuIEl0IHJpZGVzIGFsb25nIGEgdGVtcGxhdGUgc3RyYW5kIG9mIEROQSBhbmQgYXR0YWNoZXMgbmV3IG51Y2xlb3RpZGVzIHRvIHRoZSBncm93aW5nIHN0cmFuZC4gTG9vayBhdCBzdGVwIDMgYW5kIHNlZSBpZiB5b3UgY2FuIGZpbmQgc29tZXRoaW5nIHRoYXQgZml0cyB0aGF0IGRlc2NyaXB0aW9uLg==[Qq]

[q multiple_choice=”true”] In the diagram below, phase 1 is best described as

[c]IGRlbmF0dX JhdGlvbg==[Qq]

[f]IEV4Y2VsbGVudCEgRGVuYXR1cmF0aW9uIGJyZWFrcyB0aGUgaHlkcm9nZW4gYm9uZHMgaG9sZGluZyBETkEgdG9nZXRoZXIsIGJyZWFraW5nIHRoZSBETkEgaW50byB0d28gc2luZ2xlIHN0cmFuZHMu[Qq]

[c]IGFubmVhbGluZw==[Qq]

[f]IE5vLiAmIzgyMjA7QW5uZWFsaW5nJiM4MjIxOyBtZWFucyBzdGlja2luZyB0by4mIzgyMjE7IEluIHBoYXNlIDEsIG5vdGhpbmcmIzgyMTc7cyBzdGlja2luZy4gUmF0aGVyIGJvbmRzIGFyZSBiZWluZyBicm9rZW4u[Qq]

[c]IGVsb25nYXRpb24=[Qq]

[f]IE5vLiBFbG9uZ2F0aW9uIGlzIHdoYXQmIzgyMTc7cyBoYXBwZW5pbmcgaW4gc3RlcCAzLg==[Qq]

[q multiple_choice=”true”] In the diagram below, phase 2 is

[c]IGRlbmF0dXJhdGlvbg==[Qq]

[f]IE5vLiBEZW5hdHVyYXRpb24gYnJlYWtzIHRoZSBoeWRyb2dlbiBib25kcyBob2xkaW5nIEROQSB0b2dldGhlciwgYnJlYWtpbmcgdGhlIEROQSBpbnRvIHR3byBzaW5nbGUgc3RyYW5kcy4gV2hhdCB5b3Ugc2VlIGluIHBoYXNlIHR3byBhcmUgdHdvIHRoaW5ncyBzdGlja2luZyB0b2dldGhlci4=[Qq]

[c]IGFubmVh bGluZw==[Qq]

[f]IEV4Y2VsbGVudC4gJiM4MjIwO0FubmVhbGluZyYjODIyMTsgbWVhbnMgc3RpY2tpbmcgdG8uJiM4MjIxOyBJbiBwaGFzZSAyLCB0aGUgcHJpbWVyIGlzIHN0aWNraW5nIHRvIHRoZSB0ZW1wbGF0ZSBzdHJhbmRzLg==[Qq]

[c]IGVsb25nYXRpb24=[Qq]

[f]IE5vLiBFbG9uZ2F0aW9uIGlzIHdoYXQmIzgyMTc7cyBoYXBwZW5pbmcgaW4gc3RlcCAzLg==[Qq]

[q multiple_choice=”true”] In the diagram below, phase 3 is

[c]IGRlbmF0dXJhdGlvbg==[Qq]

[f]IE5vLiBEZW5hdHVyYXRpb24gYnJlYWtzIHRoZSBoeWRyb2dlbiBib25kcyBob2xkaW5nIEROQSB0b2dldGhlciwgYnJlYWtpbmcgdGhlIEROQSBpbnRvIHR3byBzaW5nbGUgc3RyYW5kcy4gV2hhdCB5b3Ugc2VlIGluIHBoYXNlIHRocmVlIGludm9sdmVzIEROQSBwb2x5bWVyYXNlIGFkZGluZyBuZXcgbnVjbGVvdGlkZXMgdGhlIEROQSBtb2xlY3VsZS4=[Qq]

[c]IGFubmVhbGluZw==[Qq]

[f]IE5vLiAmIzgyMjA7QW5uZWFsaW5nJiM4MjIxOyBtZWFucyBzdGlja2luZyB0bywmIzgyMjE7IGFuZCBpdCByZWZlcnMgdG8gdGhlIHByaW1lciBzdGlja2luZyB0byB0aGUgdGVtcGxhdGUgc3RyYW5kLg==[Qq]

[c]IGVsb25n YXRpb24=[Qq]

[f]IFdheSB0byBHbyEgRWxvbmdhdGlvbiBpcyB3aGF0JiM4MjE3O3MgaGFwcGVuaW5nIGluIHN0ZXAgMy4=[Qq]

[q] In the diagram below, which letter indicates annealing?

[textentry single_char=”true”]

[c]IG M=[Qq]

[f]WWVzISBMZXR0ZXIgJiM4MjIwO2MmIzgyMjE7IHJlcHJlc2VudHMgYW5uZWFsaW5nLg==[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLg==[Qq]

[c]ICo=[Qq]

[f]IE5vLiAmIzgyMjA7QW5uZWFsaW5nJiM4MjIxOyBtZWFucyAmIzgyMjA7c3RpY2tpbmcgdG8uJiM4MjIxOyBGaW5kIHdoZXJlIHRoZSBwcmltZXIgaXMgc3RpY2tpbmcgdG8gdGhlIEROQSB0ZW1wbGF0ZS4=[Qq]

[q] In the diagram below, which letter indicates elongation?

[textentry single_char=”true”]

[c]IG Q=[Qq]

[f]WWVzISBMZXR0ZXIgJiM4MjIwO2QmIzgyMjE7IHJlcHJlc2VudHMgZWxvbmdhdGlvbi4=[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLCB0aGF0JiM4MjE3O3Mgbm90IGNvcnJlY3Qu[Qq]

[c]ICo=[Qq]

[f]IE5vLiBGaW5kIHdoZXJlIEROQSBwb2x5bWVyYXNlIGlzIGNyZWF0aW5nIGFuIGluY3JlYXNpbmdseSBsb25nIHN0cmFuZCBvZiBjb21wbGVtZW50YXJ5IEROQS4=[Qq]

[q] In the diagram below, which number is pointing to a primer?

[textentry single_char=”true”]

[c]ID I=[Qq]

[f]TmljZSBqb2IuICYjODIyMDsyJiM4MjIxOyByZXByZXNlbnRzIGEgcHJpbWVyLg==[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IFNvcnJ5LCB0aGF0JiM4MjE3O3Mgbm90IGNvcnJlY3Qu[Qq]

[c]ICo=[Qq]

[f]IE5vLiBUaGUgcHJpbWVyIGlzIGEgc2hvcnQgc2VxdWVuY2Ugb2Ygc2luZ2xlLXN0cmFuZGVkIEROQSB0aGF0IGJpbmRzIHdpdGggdGhlIDMmIzgyNDI7IGVuZCBvZiB0aGUgdGVtcGxhdGUgRE5BLg==[Qq]

[q] In the diagram below, which number is pointing to free nucleotides?

[textentry single_char=”true”]

[c]ID M=[Qq]

[f]WWVzISAmIzgyMjA7MyYjODIyMTsgcmVwcmVzZW50cyBmcmVlIG51Y2xlb3RpZGVzLg==[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLg==[Qq]

[c]ICo=[Qq]

[f]IE5vLiBUaGUgZnJlZSBudWNsZW90aWRlcyB3aWxsIGJlIHVzZWQgZHVyaW5nIHRoZSBlbG9uZ2F0aW9uIHBoYXNlIHRvIGNyZWF0ZSBuZXcgc3RyYW5kcyBvZiBETkEu[Qq]

[q] In the diagram below, which number is pointing to newly synthesized DNA?

[textentry single_char=”true”]

[c]ID Q=[Qq]

[f]RXhjZWxsZW50LiAmIzgyMjA7NCYjODIyMTsgcmVwcmVzZW50cyBuZXdseSBzeW50aGVzaXplZCBETkEu[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLg==[Qq]

[c]ICo=[Qq]

[f]IE5vLiBUaGUgbmV3IEROQSBpcyB3aGF0JiM4MjE3O3MgYmVpbmcgc3ludGhlc2l6ZWQgZHVyaW5nIHRoZSBlbG9uZ2F0aW9uIHBoYXNlLg==[Qq]

[q] In the diagram below, which number indicates DNA polymerase?

[textentry single_char=”true”]

[c]ID U=[Qq]

[f]V2F5IHRvIGdvLiBOdW1iZXIgJiM4MjIwOzUmIzgyMjE7IHNob3dzIEROQSBwb2x5bWVyYXNlLg==[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IFNvcnJ5LCB0aGF0JiM4MjE3O3Mgbm90IGNvcnJlY3Qu[Qq]

[c]ICo=[Qq]

[f]IE5vLiBETkEgcG9seW1lcmFzZSBpcyB0aGUgZW56eW1lIHRoYXQmIzgyMTc7cyBzeW50aGVzaXppbmcgbmV3IEROQSBkdXJpbmcgdGhlIGVsb25nYXRpb24gcGhhc2Uu[Qq]

[q] The image below shows a closeup view of what’s happening during PCR. The primer must be at number

[textentry single_char=”true”]

[c]ID E=[Qq]

[f]IEV4Y2VsbGVudCEgVGhlIHByaW1lciBpcyBhdCAmIzgyMjA7MS4mIzgyMjE7[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLg==[Qq]

[c]ICo=[Qq]

[f]IE5vLiBUaGUgcHJpbWVyIGlzIGEgc2hvcnQgc3RyYW5kIG9mIEROQSB0aGF0IGJpbmRzIHdpdGggdGhlIHRlbXBsYXRlIHN0cmFuZC4=[Qq]

[q] The image below shows a closeup view of what’s happening during PCR. Newly synthesized DNA must be at number

[textentry single_char=”true”]

[c]ID M=[Qq]

[f]IEdyZWF0ISBOdW1iZXIgJiM4MjIwOzMmIzgyMjE7IGlzIHRoZSBuZXdseSBzeW50aGVzaXplZCBETkEu[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLCB0aGF0JiM4MjE3O3Mgbm90IGNvcnJlY3Qu[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50OiB0aGUgdGVtcGxhdGUgRE5BIGlzIGF0ICYjODIyMDsyLiYjODIyMTs=[Qq]

[q] The image below shows a closeup view of what’s happening during PCR. DNA polymerase must be at

[textentry single_char=”true”]

[c]ID U=[Qq]

[f]IEdvb2QhIEROQSBwb2x5bWVyYXNlIGlzIGF0IG51bWJlciA1LsKg[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLg==[Qq]

[c]ICo=[Qq]

[f]IE5vLiBETkEgcG9seW1lcmFzZSBpcyB0aGUgZW56eW1lIHRoYXQmIzgyMTc7cyBjb25uZWN0aW5nIGZyZWUgbnVjbGVvdGlkZXMgdG8gY3JlYXRlIG5ldyBETkEu[Qq]

[q] PCR stands for[hangman] [hangman] [hangman]

[c]IHBvbHltZXJhc2U=[Qq]

[c]Y2hhaW4=[Qq]

[f]IEdyZWF0IQ==[Qq]

[c]IHJlYWN0aW9u[Qq]

[f]IENvcnJlY3Qh[Qq]

[q] The image below shows a closeup view of what’s happening during PCR. Free nucleotide triphosphates are shown at
[textentry single_char=”true”]

[c]ID Q=[Qq]

[f]IEdyZWF0ISBGcmVlIG51Y2xlb3RpZGVzIGFyZSBzaG93biBhdCAmIzgyMjA7NC4mIzgyMjE7[Qq]

[c]IEVudGVyIHdvcmQ=[Qq]

[f]IE5vLCB0aGF0JiM4MjE3O3Mgbm90IGNvcnJlY3Qu[Qq]

[c]ICo=[Qq]

[f]IE5vLiBIZXJlJiM4MjE3O3MgYSBoaW50OiBmcmVlIG51Y2xlb3RpZGVzIGFyZSBnb2luZyB0byBiZSBpbmNvcnBvcmF0ZWQgaW50byB0aGUgbmV3bHkgc3ludGhlc2l6ZWQgc3RyYW5kLCB3aGljaCBpcyBhdCAmIzgyMjA7My4mIzgyMjE7IFdoYXQgY291bGQgYmUgdGhlIG1vbm9tZXJzIHRoYXQgZ2V0IGluY29ycG9yYXRlZCBpbnRvIHRoYXQgc3RyYW5kPw==[Qq]

[q] During PCR, the highest heat is applied during the[hangman]phase

[c]IGRlbmF0dXJhdGlvbg==[Qq]

[q] During PCR, primers bind to DNA during the [hangman] phase.

[c]IGFubmVhbGluZw==[Qq]

[/qwiz]

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