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Questions and Answers
What occurs when 5-methylcytosine is deaminated?
What occurs when 5-methylcytosine is deaminated?
What is a consequence of pyrimidine dimers in DNA?
What is a consequence of pyrimidine dimers in DNA?
Which statement about ATP hydrolysis is true?
Which statement about ATP hydrolysis is true?
Which enzyme synthesizes RNA?
Which enzyme synthesizes RNA?
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What role do nucleotides play as coenzymes?
What role do nucleotides play as coenzymes?
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What is a key characteristic of nanopore sequencing?
What is a key characteristic of nanopore sequencing?
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Which statement is true regarding the location of DNA in prokaryotic cells like E.coli?
Which statement is true regarding the location of DNA in prokaryotic cells like E.coli?
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How does DNA supercoiling help in cellular function?
How does DNA supercoiling help in cellular function?
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What is NOT a characteristic of nanopore sequencing?
What is NOT a characteristic of nanopore sequencing?
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What role does helicase play in nanopore sequencing?
What role does helicase play in nanopore sequencing?
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What is the primary purpose of using exonuclease in the cloning of recombinant DNA?
What is the primary purpose of using exonuclease in the cloning of recombinant DNA?
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What characteristic of the lac Z gene is crucial for identifying successful cloning in bacteria?
What characteristic of the lac Z gene is crucial for identifying successful cloning in bacteria?
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What distinguishes eukaryotic cells from prokaryotic cells regarding chromosome structure?
What distinguishes eukaryotic cells from prokaryotic cells regarding chromosome structure?
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What is the role of centromeres in yeast artificial chromosomes (YAC)?
What is the role of centromeres in yeast artificial chromosomes (YAC)?
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Which step in PCR specifically involves adding primers to the DNA template?
Which step in PCR specifically involves adding primers to the DNA template?
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What is a significant feature of reverse transcriptase PCR?
What is a significant feature of reverse transcriptase PCR?
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What is the function of a transcription terminator sequence in expression vectors?
What is the function of a transcription terminator sequence in expression vectors?
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What process results in exponential amplification of DNA during PCR?
What process results in exponential amplification of DNA during PCR?
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What is the purpose of tagging fusion proteins to a reporter gene like GFP?
What is the purpose of tagging fusion proteins to a reporter gene like GFP?
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Which method uses viral vectors to introduce DNA into cells?
Which method uses viral vectors to introduce DNA into cells?
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What is a primary characteristic of SNPs?
What is a primary characteristic of SNPs?
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What is a limitation of Sanger sequencing?
What is a limitation of Sanger sequencing?
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What composes the majority of the human genome?
What composes the majority of the human genome?
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How has the cost of sequencing the human genome changed?
How has the cost of sequencing the human genome changed?
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What is the primary function of the terminator nucleotide in Sanger sequencing?
What is the primary function of the terminator nucleotide in Sanger sequencing?
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What enhances the evolutionary rate in regulatory DNA in humans?
What enhances the evolutionary rate in regulatory DNA in humans?
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What is the function of reverse transcriptase in the PCR process?
What is the function of reverse transcriptase in the PCR process?
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What is a key characteristic of real-time PCR?
What is a key characteristic of real-time PCR?
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Which method is used for amplifying specific DNA regions in DNA fingerprinting?
Which method is used for amplifying specific DNA regions in DNA fingerprinting?
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What is the purpose of using chromatography in protein purification?
What is the purpose of using chromatography in protein purification?
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In the context of producing recombinant proteins, how are proteins tagged?
In the context of producing recombinant proteins, how are proteins tagged?
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Why is it important to purify proteins in medical applications?
Why is it important to purify proteins in medical applications?
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What is the mechanism of column chromatography in protein purification?
What is the mechanism of column chromatography in protein purification?
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What is the primary role of antibodies in medical treatments?
What is the primary role of antibodies in medical treatments?
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Study Notes
DNA Damage and Repair
- Uracil is not typically found in DNA, making its presence easy to recognize.
- Deamination of 5-methylcytosine transforms it to thymine, creating a repair challenge since thymine is naturally a component of DNA.
- Pyrimidine dimers, caused by UV light, link two thymine or cytosine bases, disrupting helical structure and preventing DNA replication.
- DNA affected by pyrimidine dimers may become trapped in the nucleus, leading to cell death.
Nucleotide Functions
- ATP hydrolysis (ATP → ADP + phosphate) is the primary method for energy release in cellular processes.
- Coenzyme A transports two-carbon units in the citric acid cycle, contributing to ATP synthesis.
- Cyclic AMP has phosphate groups at both 3' and 5' positions.
- Notable nucleotide coenzymes:
- ATP provides energy for biochemical reactions.
- NAD+ is crucial for energy transfer within cells.
DNA Technologies
- DNA and RNA polymerases synthesize DNA and RNA, respectively.
- Cloning produced by transformed cells yields abundant recombinant DNA without needing restriction enzymes.
- Artificial BAC (Bacterial Artificial Chromosomes) enable cloning large DNA sequences and rely on lac Z for selection through color changes.
- Eukaryotic chromosomes are linear, larger than prokaryotic circular chromosomes, with multiple origins of replication.
PCR (Polymerase Chain Reaction)
- Denaturation, annealing with primers, and extension using Taq polymerase are the key steps in PCR.
- Variants include:
- Reverse Transcriptase PCR converts mRNA to cDNA.
- Quantitative PCR measures DNA concentrations in real time, using fluorescent dyes.
- Directed mutagenesis introduces specific changes via non-complementary primers.
- DNA fingerprinting amplifies Short Tandem Repeats (STR) for identification.
Laboratory Analysis of DNA
- Gel electrophoresis separates DNA fragments by size and charge, exploiting DNA's negative charge.
- Recombinant proteins can be produced using proteins instead of animal sources, which is important for therapeutic applications like insulin production.
Protein Purification
- Proteins are purified based on properties such as charge and size, often through chromatography techniques.
- Fusion proteins, with reporter genes (like GFP for visibility under UV light), facilitate purification processes.
Transgenic Organisms and Gene Therapy
- Gene modification technologies include electroporation, microinjection, liposomes, and viral vectors for effective gene transfer.
Human Genome Insights
- Sequencing human genomes is now affordable, identifying potential disease risks associated with individual lifestyles.
- A small portion of the human genome codes for proteins, with many regions consisting of introns.
SNPs and Haplotype Analysis
- Single nucleotide polymorphisms (SNPs) are common variations among humans.
- Haplotypes indicate specific combinations of SNPs that can distinguish individuals and disease susceptibility.
Nucleic Acid Sequencing Techniques
- Sanger sequencing uses terminator nucleotides for controlled sequencing but is limited in scale.
- Nanopore sequencing employs electrical current to directly read DNA as it passes through a nanopore, allowing simultaneous sequencing of millions of strands.
DNA Structure in Organisms
- E. coli, as a prokaryote, lacks a nucleus and organizes its genetic material into a compact nucleoid structure.
- Supercoiling is a mechanism for compacting DNA, akin to a twisted phone cord secured at both ends.
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Description
Test your knowledge on DNA damage, focusing on issues like uracil in DNA and thymine formation from deaminated methylcytosine. Additionally, explore the effects of UV light on DNA, particularly the formation of pyrimidine dimers. This quiz will cover various aspects of DNA stability and repair mechanisms.
