DNA Sequencing and Cloning Techniques
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Questions and Answers

What technique can provide clues to the function of a gene of unknown function?

  • RNA sequencing
  • In situ hybridization
  • DNA microarray analysis
  • Experimental gene knockout (correct)

How are induced pluripotent stem (iPS) cells generated?

  • By directly isolating embryonic stem cells
  • By reprogramming differentiated cells (correct)
  • By transplanting nuclei into enucleated eggs
  • By cloning an entire organism

What is a potential use of personalized medicine?

  • Increasing gene mutation rates
  • Promoting genetic disorders
  • Minimizing known risks for diseases (correct)
  • Standardized treatment for all patients

Which method can specifically edit genes in living cells?

<p>CRISPR-Cas9 system (B)</p> Signup and view all the answers

What kind of cells can give rise to any tissue in a new plant?

<p>Totipotent cells (C)</p> Signup and view all the answers

What is a key feature of genome-wide association studies (GWAS)?

<p>They utilize single nucleotide polymorphisms (SNPs) as markers (A)</p> Signup and view all the answers

What advantage do transgenic organisms offer in agriculture?

<p>Improvement in agricultural productivity (A)</p> Signup and view all the answers

What is the primary function of DNA ligase in the process of creating recombinant DNA molecules?

<p>To seal base-paired fragments of DNA (D)</p> Signup and view all the answers

What is one application of DNA technology in forensic science?

<p>Analyzing short tandem repeats (STRs) for genetic profiling (B)</p> Signup and view all the answers

Which type of stem cell is difficult to acquire yet pluripotent?

<p>Embryonic stem cells (B)</p> Signup and view all the answers

Which method is primarily used for amplification of a specific target segment of DNA?

<p>Polymerase chain reaction (PCR) (D)</p> Signup and view all the answers

What is a purpose of using genetic profiles in criminal investigations?

<p>To definitively prove a suspect's guilt or innocence (C)</p> Signup and view all the answers

What is the role of restriction enzymes in DNA cloning?

<p>To cut DNA at specific nucleotide sequences (A)</p> Signup and view all the answers

How do next-generation sequencing techniques primarily determine the sequence of nucleotides?

<p>By tracking the order in which nucleotides are added during synthesis (A)</p> Signup and view all the answers

What is a significant challenge when expressing cloned eukaryotic genes in bacterial host cells?

<p>Differences in post-translational modification of proteins (B)</p> Signup and view all the answers

What characteristic of sticky ends facilitates the process of DNA cloning?

<p>They can form hydrogen bonds with complementary sequences (D)</p> Signup and view all the answers

Which of the following statements about dideoxy sequencing is true?

<p>It adds modified nucleotides that terminate DNA strand extension (C)</p> Signup and view all the answers

What technique is used to separate DNA restriction fragments based on size?

<p>Gel electrophoresis (C)</p> Signup and view all the answers

What is the key advantage of using cultured eukaryotic cells as host cells for gene expression?

<p>They can perform necessary post-translational modifications. (C)</p> Signup and view all the answers

What does nucleic acid hybridization rely on for the detection of specific mRNAs?

<p>The pairing of nucleotides with complementary sequences (A)</p> Signup and view all the answers

Flashcards

DNA sequencing

Determining the order of nucleotides in a DNA molecule.

Dideoxy sequencing

A method for DNA sequencing using chain-terminating dideoxynucleotides.

Next-generation sequencing

High-throughput DNA sequencing techniques based on sequencing by synthesis.

Gene cloning

Making multiple copies of a gene or DNA segment.

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Restriction enzymes

Bacterial enzymes that cut DNA at specific sequences.

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Recombinant DNA

DNA molecule formed by combining DNA from different sources.

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Gel electrophoresis

Separating DNA fragments of different lengths.

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Polymerase Chain Reaction (PCR)

A technique to amplify a specific DNA segment.

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Nucleic acid hybridization

Base pairing between complementary strands of nucleic acids.

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Expression vectors

Vectors used to express a cloned eukaryotic gene in host cells.

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mRNA detection methods

Methods like in situ hybridization and RT-PCR detect the presence of specific messenger RNA (mRNA) in tissues or RNA samples.

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RNA-seq

RNA sequencing, or RNA-seq, is used to find sets of genes that are expressed together in a group of cells.

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Gene Knockout

Experimental inactivation of a gene to understand its function by observing the resulting changes in the organism.

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CRISPR-Cas9

A gene-editing tool allowing specific changes in genes of living cells.

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Gene drive

Altered alleles inherited preferentially through a population.

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Genome-wide association studies (GWAS)

GWAS use SNPs to find gene variations linked to specific conditions in humans.

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Totipotent cells

Plant cells able to create all plant tissues.

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Organismal cloning

Creating a new organism with the same genetic makeup as an existing one, often using nuclear transfer.

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Gene therapy factors

Factors that decide if a genetic illness is suitable for effective gene therapy.

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iPS cells

A type of stem cell created by reprogramming differentiated adult cells, similar in function to embryonic stem cells.

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Study Notes

DNA Sequencing and Cloning

  • DNA sequencing methods include dideoxy sequencing, next-generation (high-throughput) sequencing by synthesis, and third-generation sequencing (e.g., nanopore).
  • Nucleic acid hybridization involves base pairing of nucleic acid strands.
  • Gene cloning creates multiple copies of a gene or DNA segment.
  • Restriction enzymes cut DNA at specific sequences (restriction sites).
  • DNA ligase joins DNA fragments.
  • Gel electrophoresis separates DNA fragments based on size.
  • Polymerase Chain Reaction (PCR) amplifies DNA segments.
  • Recombinant plasmids are introduced into host cells, creating clones.
  • Eukaryotic gene expression is challenging in bacteria – eukaryotic cells are a better host.
  • Expression vectors are essential for eukaryotic gene expression.

Using DNA Technology to Study Gene Expression

  • Nucleic acid probes detect specific mRNAs.
  • In situ hybridization and RT-PCR locate mRNA in tissues and samples.
  • RNA-seq and DNA microarrays analyze sets of co-expressed genes.
  • Gene knockouts identify gene function through observation of phenotypic effects.
  • CRISPR-Cas9 allows precise gene editing.
  • Genome-wide association studies identify genetic markers (SNPs) associated with diseases.

Applying Cloned Organisms and Stem Cells

  • Totipotent plant cells can generate complete plants.
  • Animal cloning involves transplanting a nucleus into an enucleated egg.
  • Embryonic stem cells (ES cells) and adult stem cells can differentiate.
  • ES cells are pluripotent.
  • Induced pluripotent stem cells (iPS cells) are similar to ES cells, more accessible– reprogrammed differentiated cells.

Practical Applications of DNA-Based Biotechnology

  • DNA technology aids in disease diagnosis, personal genome analysis, and personalized medicine.
  • Gene therapy aims to cure genetic disorders or cancer.
  • Therapeutic proteins are created using cell cultures and transgenic animals.
  • DNA analysis aids in crime investigations, paternity disputes, and victim identification.
  • Genetically engineered microorganisms can extract minerals or degrade waste.
  • Transgenic plants and animals aim to enhance agricultural productivity and food quality.
  • Benefits of genetic engineering must be weighed against risks to humans and the environment.

Gene Cloning in Cells

  • Recombinant plasmids contain foreign DNA.
  • Plasmids are inserted into bacterial cells.
  • Cloning produces cells with identical genetic information.
  • Cells multiply and produce many copies of the plasmid.

Choosing Candidates for Gene Therapy

  • Factors to consider for successful gene therapy: disease severity, accessibility of target cells, and potential for harmful side effects.
  • The safety and efficacy of the gene therapy method.

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Description

This quiz covers essential methods in DNA sequencing and cloning, including various sequencing techniques, gene cloning processes, and key biochemical tools like PCR and gel electrophoresis. Additionally, explore the challenges and technologies related to gene expression in eukaryotic systems. Test your understanding of these fundamental concepts in molecular biology.

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