Microbiology 329 - Chromosomal gDNA Extractions and GASP Competition Assay
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These flashcards cover key concepts related to chromosomal gDNA extractions and the setup of GASP competition assays in microbiology.
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13 Terms
Chromosomal gDNA Extractions
The process of isolating genomic DNA from cells, involving steps like cell lysis, protein precipitation, and DNA precipitation.
Cell Lysis
The process of breaking open cells to release their contents, including DNA, through various methods such as mechanical, chemical, or enzymatic.
Protein Precipitation
The process of removing proteins from a solution, often achieved by adding salts to 'salt out' the proteins.
DNA Precipitation
The process of isolating DNA from a solution by adding alcohol, which reduces the solubility of DNA and causes it to aggregate and precipitate.
Alkaline Phosphatase
An enzyme that removes phosphate groups from the ends of DNA strands, thus playing a role in DNA processing.
Ammonium Sulfate
A chemical used in protein precipitation due to its high solubility in water and ability to increase ionic strength.
GASP mutant
A genetically modified organism (mutant) being tested for relative fitness in comparison to a wild-type organism during competition assays.
1:1000 ratio
The dilution ratio of the GASP mutant to wild-type cells, ensuring that the mutant makes up 0.1% of the population to demonstrate its relative fitness.
Relative Fitness
A measure of how well a genotype performs in comparison to others in a specific environment, often evaluated through competition assays.
Isopropanol
An alcohol used in DNA precipitation to help the DNA come out of solution.
Phenol:Chloroform:Isoamyl Alcohol
A traditional method for DNA extraction that utilizes a mixture of solvents to separate DNA from proteins and other cellular components.
Sterile Water Storage
Short-term storage solution for DNA, adequate for temporary preservation.
Buffered Solution for DNA Stability
A long-term storage solution, such as 10 mM Tris-Cl, pH 8.5, that helps maintain the structural integrity of DNA.
