Genetics Week 8 Quiz

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Questions and Answers

What technique is used to separate DNA fragments into different sizes?

PCR

Molecular Cloning

Southern blotting

Gel Electrophoresis (correct)

Which technique would you use to isolate a specific gene of interest after separating DNA fragments?

Gel Electrophoresis

Southern blotting (correct)

DNA Sequencing

Molecular Cloning

Why is SDS used in SDS-PAGE?

To increase the pH of the solution

To provide nutrients to the gel

To enhance the enzyme activity

To denature proteins and provide a uniform charge (correct)

What is the role of primary antibodies in Western blotting?

To bind to the target protein Signup and view all the answers

Which chemical is commonly used to stain protein fragments on an SDS-PAGE gel?

Coomassie Blue Signup and view all the answers

After obtaining large quantities of a gene, which technique is used to determine the DNA sequence?

DNA Sequencing Signup and view all the answers

What is a characteristic of Nitrocellulose membranes used in Western blotting?

They have a high protein-binding capacity Signup and view all the answers

What technique would you use to clone and produce a large quantity of a gene?

Molecular Cloning Signup and view all the answers

What is used to separate proteins based on size in SDS-PAGE?

Electric current Signup and view all the answers

Which technique is applied to express protein products from the isolated gene?

Recombinant Protein Expression Signup and view all the answers

What is the primary function of primary antibodies in a western blot?

To bind specifically to the target protein Signup and view all the answers

What is the role of secondary antibodies in western blotting?

To bind to the primary antibody and enable detection Signup and view all the answers

What differentiates dideoxynucleotide triphosphates (ddNTPs) from deoxynucleotide triphosphates (dNTPs) in Sanger sequencing?

ddNTPs lack a 3’ hydroxyl group Signup and view all the answers

Which technique is used as a confirmatory test for HIV through western blotting?

Detection of anti-HIV antibodies Signup and view all the answers

What does the term 'genomics' refer to?

The complete set of DNA in an organism Signup and view all the answers

What is the initial step in Hierarchical Shotgun Sequencing?

Cloning DNA into bacterial artificial chromosomes (BACs) Signup and view all the answers

Which of the following is true about SNPs?

They are the most common type of genetic variation Signup and view all the answers

What is the main function of Short Tandem Repeat (STR) Analysis in DNA profiling?

To examine unique DNA sequences based on repeat variations Signup and view all the answers

What is a haplotype?

A group of alleles inherited together on a single chromosome Signup and view all the answers

In which DNA profiling technique do restriction enzymes play a critical role?

Restriction Fragment Length Polymorphism (RFLP) Signup and view all the answers

Study Notes

Molecular Techniques for DNA and Protein Analysis

Gel Electrophoresis: Method for separating DNA fragments by size after isolation from an organism.
Southern Blotting: Used to isolate specific genes of interest post-separation of DNA fragments.
Molecular Cloning: Technique to clone and produce large quantities of a targeted gene.
Reverse Transcription (RT) & CR: Processes for understanding transcriptional profiles of genes, measuring gene expression levels.
DNA Sequencing: Determines the precise sequence of nucleotides in DNA, checking for mutations.
Recombinant Protein Expression: Produces protein products from cloned genes.
SDS-PAGE: Technique for separating proteins based on size after tissue collection.
Western Blotting: Isolates and detects specific proteins post-SDS-PAGE separation.

SDS-PAGE and Protein Visualization

Role of SDS: Sodium Dodecyl Sulfate denatures proteins by unfolding them and imparts a uniform negative charge, allowing size-based separation during electrophoresis.
Staining Agents: Coomassie Blue is commonly used to visualize protein bands on SDS-PAGE gels.
Membrane Types: Nitrocellulose and Polyvinylidene Difluoride (PVDF) membranes are commonly used in Western blotting.
Protein Detection: Uses antibodies; primary antibodies bind to target proteins, while secondary antibodies facilitate detection via conjugation with enzymes or fluorophores.
Antibody Functions:
- Primary antibodies specifically bind to target proteins.
- Secondary antibodies bind to primary antibodies to enable visualization or detection of target proteins.
Western Blot Clinical Application: Used for confirming HIV diagnosis by detecting anti-HIV antibodies in serum.

Sanger Sequencing

Modified Nucleotides: Dideoxynucleotide triphosphates (ddNTPs) lack a 3’ hydroxyl group, preventing chain elongation when incorporated into the DNA strand, thus terminating synthesis.

Genomics Overview

Definition: Genomics is the comprehensive study of an organism’s complete DNA (genome), including structure, function, evolution, and mapping.

Human Genome Project Sequencing Methods

Hierarchical Shotgun Sequencing: Large DNA fragments are cloned, mapped, and sequenced in smaller pieces to create a physical map.
Whole Genome Shotgun Sequencing: Entire genomes are fragmented and sequenced directly, assembling sequences using computational methods.

Genetic Variations

Single Nucleotide Polymorphisms (SNPs): Variations at a single nucleotide position in DNA, representing common genetic variations among individuals.
Haplotype: A specific combination of alleles or SNPs on a single chromosome that are inherited together.

DNA Profiling Techniques

Short Tandem Repeat (STR) Analysis: Examines repeating base sequences in DNA, varying among individuals, useful for identification.
Restriction Fragment Length Polymorphism (RFLP): Cuts DNA into fragments using restriction enzymes and separates them by gel electrophoresis, revealing individual variations.

Southern Blot vs. Western Blot

Hybridization Probes in Southern Blot: Used to detect specific DNA sequences by binding to complementary strands.
Western Blot Techniques: Utilizes antibodies for the detection of specific proteins.

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Description

This quiz covers the molecular techniques used in genetic analysis, focusing on methods such as gel electrophoresis, southern blotting, and molecular cloning. It encourages students to explore and apply these techniques in various scenarios related to DNA manipulation. Test your understanding of genetics and the methodologies for analyzing DNA fragments!