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Questions and Answers
What is the primary function of reverse transcriptase?
What is the primary function of reverse transcriptase?
Which type of DNA is synthesized from RNA transcripts?
Which type of DNA is synthesized from RNA transcripts?
In gel electrophoresis, what factors determine the rate at which DNA fragments move through the gel?
In gel electrophoresis, what factors determine the rate at which DNA fragments move through the gel?
What information can be obtained from the patterns produced in gel electrophoresis?
What information can be obtained from the patterns produced in gel electrophoresis?
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What is a significant use of gene probes?
What is a significant use of gene probes?
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What is the primary purpose of fluorescent in situ hybridization (FISH)?
What is the primary purpose of fluorescent in situ hybridization (FISH)?
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Which component is NOT involved in the Polymerase Chain Reaction (PCR)?
Which component is NOT involved in the Polymerase Chain Reaction (PCR)?
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How does nucleic acid hybridization detect the presence of specific sequences?
How does nucleic acid hybridization detect the presence of specific sequences?
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Which DNA size is equivalent to the length of an average gene in E. coli?
Which DNA size is equivalent to the length of an average gene in E. coli?
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What is a key feature of PCR that distinguishes it from other DNA replication techniques?
What is a key feature of PCR that distinguishes it from other DNA replication techniques?
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Which of the following is NOT a benefit of using FISH?
Which of the following is NOT a benefit of using FISH?
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What role do primers play in the Polymerase Chain Reaction (PCR)?
What role do primers play in the Polymerase Chain Reaction (PCR)?
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What is the estimated size of the human genome in base pairs?
What is the estimated size of the human genome in base pairs?
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What is the first step of the PCR technique?
What is the first step of the PCR technique?
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Which method is used to detect products during the reaction in real-time PCR?
Which method is used to detect products during the reaction in real-time PCR?
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What is required for RNA to be analyzed using PCR?
What is required for RNA to be analyzed using PCR?
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In recombinant DNA technology, what is the primary purpose of a vector?
In recombinant DNA technology, what is the primary purpose of a vector?
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Which aspect is crucial for a cloning vector to function properly?
Which aspect is crucial for a cloning vector to function properly?
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What is the role of plasmids in cloning vectors?
What is the role of plasmids in cloning vectors?
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Which of the following best describes genomic libraries?
Which of the following best describes genomic libraries?
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What is a characteristic of bacteriophage used as cloning vectors?
What is a characteristic of bacteriophage used as cloning vectors?
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What role do restriction endonucleases play in genetic engineering?
What role do restriction endonucleases play in genetic engineering?
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Which of the following describes 'sticky ends' in genetic engineering?
Which of the following describes 'sticky ends' in genetic engineering?
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What is the main application of ligase in genetic engineering?
What is the main application of ligase in genetic engineering?
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What are restriction fragments?
What are restriction fragments?
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How do restriction fragment length polymorphisms (RFLPs) aid in genetic comparison?
How do restriction fragment length polymorphisms (RFLPs) aid in genetic comparison?
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Which statement regarding the application of genetic engineering in agriculture is correct?
Which statement regarding the application of genetic engineering in agriculture is correct?
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What does the term 'applied science' refer to in the context of genetic engineering?
What does the term 'applied science' refer to in the context of genetic engineering?
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What is the significance of foreign DNA recognition by restriction endonucleases?
What is the significance of foreign DNA recognition by restriction endonucleases?
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Study Notes
Introduction to Genetic Engineering
- Basic science: Focuses on fundamental processes, no direct product or application arises
- Example: microorganism duplication, exchange, and genetic information usage
Applied Science
- Applications derived from basic research
- Examples:
- Genetic manipulation to select only advantageous genes in pigs
- Using DNA to identify suspects in crimes
- Fixing genetic mutations to treat diseases
- Utilizing RNA regulatory molecules to permanently "fix" diseases
DNA: The Raw Material
- Intrinsic properties:
- Helicase unwinds double helix as easily in lab as in a cell
- DNA strands separate at temperatures just below boiling (96 °C)
- Complementary strands re-form when cooled
Enzymes for Splicing, Dicing, and Reversing Nucleic Acids
- Restriction endonucleases:
- Cut DNA crosswise at specific sites
- Recognize foreign DNA
- Break phosphodiester bonds between nucleotides in DNA
- Purpose: protect bacteria from bacteriophages or plasmids
- Recognition of palindromes (specific sequences that read the same forward and backward)
- Production of sticky ends (short, single-stranded overhangs) making easy incorporation of DNA fragments possible
Sticky Ends
- Staggered cuts, leaving tails of 4-5 bases
- Base pairs with complementary tails on other DNA fragments or plasmids
Ligase
- Necessary to seal sticky ends together
- Joins phosphate-sugar bonds cut by endonucleases
- Used to splice genes into plasmids and chromosomes
Restriction Fragments
- Pieces of DNA produced by restriction endonucleases
- Restriction Fragment Length Polymorphisms (RFLPs):
- Differences in specific restriction endonucleases' cutting patterns create variations in fragment lengths
Reverse Transcriptase and Complementary DNA (cDNA)
- Reverse transcriptase:
- Replicates HIV and other retroviruses
- Converts RNA into DNA
- cDNA:
- Allows direct comparison of DNA from different organisms at a specific site.
- Made from messenger, transfer, ribosomal, and other forms of RNA
- Useful in synthesizing eukaryotic genes from mRNA transcripts
What are Genes?
- Genes are located on a DNA molecule within a chromosome within a cell.
What are Introns?
- Introns are sections of a gene that are not included in the final mRNA molecule.
- They do not code for amino acids.
Exons vs. Introns
- Exons are coding sections of a gene; Introns are non-coding sections
- Exons comprise the final mRNA; Introns are removed during processing.
Analysis of DNA
- Gel electrophoresis:
- Creates a pattern of DNA fragments for analysis
- DNA migrates through a gel based on fragment size.
- Positions determined by staining.
- Useful in characterizing DNA fragments, and comparing genetic similarities.
- RFLP (Restriction Fragment Length Polymorphism):
- Used for comparison of DNA.
- Different lengths of restriction fragments show genetic difference between individuals.
- Nucleic Acid Hybridization:
- Two different nucleic acids can unite at their complementary regions
- Gene Probes:
- Short stretches of DNA to find complementary DNA sequences.
- Carry reporter molecules (e.g., fluorescent dyes) for visualization during hybridization
Polymerase Chain Reaction (PCR)
- A molecular xerox machine, rapidly increasing DNA copies
- Highly sensitive (detects single cells)
- PCR uses similar events as normal DNA replication. specialized ingredients include primers and DNA polymerases
- PCR technique includes the use of a thermal cycler; basic steps include denaturation, priming, and extension.
- Real time PCR detects products during reaction, and PCR can be adapted to analyze RNA
Methods in Recombinant DNA Technology
- Recombinant DNA technology:
- Deliberate removal of genetic material from one organism to combine with another.
- Creation of identical copies of DNA
- Cloning:
- Removes a selected gene from an organism (genetic donor).
- Propagation of that gene in a different host organism.
- Cloning Vectors:
- Plasmids and bacteriophages have the capacity to accept DNA fragments of desired size.
- Plasmids usually contain a gene for drug resistance to the host cell.
- Cloning Hosts:
- Organisms chosen for cloning should have a fast growth rate, be easily cultured, not pathogenic, well-characterized genomes, and be able to accept vectors into their cells
Genetic Treatments: Introducing DNA into the Body
- Gene therapy:
- Normal gene inserted to correct a faulty gene.
- Benefit: permanent cure of physical dysfunction.
- Strategies for gene therapy:
- Insert normal gene into retrovirus or adenovirus vectors.
- Incubate patient's tissues with modified viruses to transfect
- Reintroduce transfected cells into patient's body.
- Naked DNA or a viral vector inserted directly into patient's tissues
- Experimentation and clinical trials of gene testing:
- Conducted on patients with genetic conditions;
- Targeted at cancer, single-gene defects, and infections;
- Early trials had significant implications
- Germline gene therapy:
- Inserts normal genes into egg, sperm, or early embryo,
- Creates heritable changes.
DNA Technology as Genetic Medicine
- Diseases from inappropriate protein expression: e.g., Alzheimer's, viral diseases, cancers
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Description
Test your knowledge on the fundamentals of genetic engineering in this quiz. Explore concepts from basic science to applied applications, including DNA characteristics and enzyme functions. Perfect for students and enthusiasts of biology and biotechnology.
