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Molecular Biology and Genomics
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1
Question
What is recombinant DNA technology?
Page 1
Answer
Recombinant DNA technology is a method that uses enzymes to cut and paste DNA from different sources, allowing the creation of new DNA molecules with desired genes.
2
Question
Define a cloning vector.
Page 4
Answer
A cloning vector is a DNA molecule used to carry foreign DNA into a host cell, where it can be replicated and expressed.
3
Question
What are plasmids?
Page 4
Answer
Plasmids are small, circular DNA molecules found in bacteria that can replicate independently of the chromosomal DNA and are commonly used as cloning vectors.
4
Question
Describe bacteriophage vectors.
Page 4
Answer
Bacteriophage vectors are derived from viruses that infect bacteria, used for cloning large DNA fragments up to 20 kb, with linear DNA structure.
5
Question
What are cosmids?
Page 4
Answer
Cosmids are hybrid vectors combining plasmid and phage features, capable of cloning DNA fragments up to 45 kb, suitable for large inserts.
6
Question
Explain expression vectors.
Page 4
Answer
Expression vectors contain regulatory sequences like promoters to drive the transcription and translation of inserted genes into proteins in host cells.
7
Question
What is a genomic library?
Page 5
Answer
A genomic library is a collection of recombinant DNA clones representing the entire genome of an organism, including both coding and non-coding regions.
8
Question
Differentiate between genomic and cDNA libraries.
Page 6
Answer
Genomic libraries contain fragments of the entire genomic DNA, while cDNA libraries are made from mRNA reverse-transcribed into DNA, representing only expressed genes.
9
Question
What is cDNA synthesis?
Page 6
Answer
cDNA synthesis involves reverse transcription of mRNA into complementary DNA using reverse transcriptase enzyme, creating a DNA copy of expressed genes.
10
Question
Describe colony hybridization.
Page 7
Answer
Colony hybridization is a screening method where bacterial colonies containing recombinant DNA are transferred to a filter and probed with labeled DNA to detect specific clones.
11
Question
What is a DNA probe?
Page 7
Answer
A DNA probe is a labeled single-stranded DNA fragment that hybridizes to complementary sequences in a target sample to detect specific genes.
12
Question
Explain the polymerase chain reaction (PCR).
Page 8
Answer
PCR is a technique to amplify specific DNA segments using primers, DNA polymerase, and thermal cycling through denaturation, annealing, and extension steps.
13
Question
What are the three main steps of PCR?
Page 8
Answer
The three steps are: 1) Denaturation (separate strands at high temperature), 2) Annealing (primers bind to target), 3) Extension (polymerase synthesizes new strands).
14
Question
What is Taq polymerase?
Page 8
Answer
Taq polymerase is a heat-stable DNA polymerase from Thermus aquaticus, used in PCR to withstand high denaturation temperatures without denaturing.
15
Question
List applications of PCR.
Page 10
Answer
Applications include DNA cloning, gene expression analysis (RT-PCR, qPCR), forensic DNA analysis, paternity testing, and diagnostic tests for diseases.
16
Question
What is gel electrophoresis?
Page 11
Answer
Gel electrophoresis separates DNA fragments by size using an electric field through a gel matrix, where smaller fragments migrate faster.
17
Question
Describe restriction mapping.
Page 12
Answer
Restriction mapping determines the locations of restriction enzyme cut sites in DNA by analyzing fragment sizes produced by different enzyme digests.
18
Question
What is Sanger sequencing?
Page 13
Answer
Sanger sequencing uses dideoxynucleotides to terminate DNA synthesis at specific bases, producing fragments separated by size to read the sequence.
19
Question
Explain next-generation sequencing (NGS).
Page 16
Answer
NGS enables massively parallel sequencing of millions of DNA fragments simultaneously, providing high-throughput, cost-effective genome analysis.
20
Question
What is the Human Genome Project (HGP)?
Page 27
Answer
The HGP was an international effort to sequence the entire human genome, completed in 2003, identifying about 20,000-25,000 genes.
21
Question
What is bioinformatics?
Page 24
Answer
Bioinformatics is the use of computational tools to analyze and interpret biological data, especially genomic sequences.
22
Question
Define Southern blotting.
Page 18
Answer
Southern blotting detects specific DNA sequences by separating fragments via electrophoresis, transferring to a membrane, and hybridizing with a probe.
23
Question
What is Northern blotting?
Page 20
Answer
Northern blotting analyzes RNA expression by separating RNA via electrophoresis, transferring to a membrane, and probing for specific transcripts.
24
Question
Explain Western blotting.
Page 20
Answer
Western blotting detects specific proteins by separating them via electrophoresis, transferring to a membrane, and probing with antibodies.
25
Question
What is fluorescence in situ hybridization (FISH)?
Page 18
Answer
FISH uses fluorescent probes to bind and visualize specific DNA sequences on chromosomes in their native cellular context.
26
Question
Describe real-time PCR (qPCR).
Page 19
Answer
Real-time PCR quantifies DNA amplification in real time using fluorescent dyes or probes, allowing measurement of gene expression levels.
27
Question
What is a microarray?
Page 22
Answer
A microarray is a high-throughput tool with thousands of DNA spots on a chip to measure gene expression or detect variations simultaneously.
28
Question
Define RNA interference (RNAi).
Page 23
Answer
RNAi is a process where small RNAs silence gene expression by degrading target mRNA or inhibiting translation.
29
Question
What are siRNAs?
Page 23
Answer
Small interfering RNAs (siRNAs) are double-stranded RNAs (21-23 nt) that trigger mRNA cleavage in the RNAi pathway.
30
Question
Explain genomics.
Page 1
Answer
Genomics is the study of the structure, function, evolution, and mapping of genomes.