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29 Cards in this Set
- Front
- Back
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What process enables to separate genetic or protein content based on mass? |
Gel electrophoresis |
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What do restriction enzymes do? |
Scan DNA and make a cut in each of the two sugar backbones of the DNA at a predefined sequence |
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What is southern blot used for? |
Visualize a specific piece of DNA of interest (e.g. a gene) to check if it is present in a sample |
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What two processes are used in southern blot? |
Restriction digestion |
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What is the DNA insert? |
A DNA piece of interest |
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Name 5 host-vector systems used in DNA cloning |
Plasmid-Bacteria Bacteriophage-Bacteria Cosmid-Bacteria Bacterial artificial chromosome-Bacteria Yeast artificial chromosome-Yeast |
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What do DNA libraries provide |
DNA libraries are collection of DNA fragments stored in large quantities by DNA cloning. |
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What is the goal of Polymerase chain reaction (PCR)? |
Replicate multiple copies of (i.e. Amplify) a segment of DNA of interest |
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Outline the process of PCR |
1. Heat dsDNA ➙ 2 ssDNA (denaturation) 2. Annealing: primers attach to ssDNA to delimit the segment of interest. 3. In presence of nucleotides and a polymerase, DNA replicates (elongation) |
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Name the ingredients needed for PCR |
- Nucleotides - Heat-resistant polymerase (Taq) - Heat - Primers |
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What is the main drawback of PCR compared to in-vivo cloning with plasmid-bacteria system? |
The nucleotide sequence of interest must be known in advance to manufacture primers |
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What technique can be used to detect RNA expression? |
Reverse transcriptase PCR |
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What is the goal of microarray analysis? |
Test whether a particular gene is expressed in a particular tissue/subject/condition. |
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What can be used to test whether a particular gene is expressed in a particular tissue/subject/condition? |
Microarray analysis |
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Outline the process of microarray analysis |
1. Microarray contains various template DNA to be tested 2. mRNA extracted from cells 3. cDNA created from mRNA by RT 4. cDNA fluorescently labeled 5. cDNA added to each spot of the array 6. Fluorescence microscope detects activated spots |
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What is the goal of sequencing? |
Identify the sequence of nucleotides in a DNA molecule of interest |
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What can be done to identify the sequence of nucleotides in a DNA molecule of interest? |
Sanger sequencing Next generation sequencing |
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Define recombination fraction |
#Recombinants/#Meioses |
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Define a centiMorgan |
Genetic distance between two loci on the genome: 1 cM is equivalent to a 1% chance of recombination |
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Name 4 types of polymorphic markers |
RFLP VNTR - Microsatellite VNTR - Minisatellite SNP |
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Compare polymorphic markers in terms of number of loci. |
RFLP > 0.1M Microsatellite > 0.1M Minisatellite > 0.01M SNP > 4M |
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Compare polymorphic markers in terms of polymorphism. |
RFLP - Moderate Microsatellite - High Minisatellite - High SNP - Moderate |
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What is the main advantage of VNTR compared to other polymorphic markers? |
They are highly polymorphic |
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What is the main advantage of SNP compared to other polymorphic markers? |
There are lots of them (hence higher resolution of genetic location) |
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Where on the genome do polymorphic markers mostly occur? |
In non-coding sequence |
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What is the goal of analysis with microsatellite? |
Identify where about in the genome the gene responsible for a disease/condition may be found |
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Name two applications of analysis with microsatellite |
Paternity test Forensics |
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What is the goal of genome-wise association studies? |
Identify the genes involved in multifactorial diseases (i.e. complex diseases whose aetiology depends on several genes) |
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What can be used to identify the genes involved in multifactorial diseases ? |
GWAS |
