04- DNA purification.pdf

Flashcards for vocabulary related to molecular cloning, DNA purification, and quantification techniques discussed in lecture.

Molecular cloning

Set of experimental techniques used to generate a population of organisms carrying the same molecule of recombinant DNA.

Recombinant DNA

DNA molecules formed by laboratory methods of genetic recombination (such as molecular cloning) to bring together genetic material from multiple sources, creating sequences that would not otherwise be found in the genome.

Restriction enzyme

An enzyme that cuts DNA at or near specific recognition nucleotide sequences known as restriction sites.

Gateway cloning

A molecular biology method that allows the transfer of DNA fragments between different DNA molecules, using site-specific recombination.

Gibson assembly

A molecular cloning method that allows joining of multiple DNA fragments in a single isothermal reaction.

Golden Gate cloning

A molecular cloning method that allows directional assembly of multiple DNA fragments using type IIS restriction enzymes and DNA ligase.

TOPO cloning

A molecular cloning method that relies on the enzyme topoisomerase I to insert DNA fragments into a plasmid vector.

DNA purification

The process of separating DNA from other cellular components such as proteins, RNA, and lipids.

DNA quantification

The process of measuring the concentration of DNA in a sample.

Lysis

The process of breaking cells open to release their contents, including nucleic acids.

Lysis buffer

A solution used to break open cells, typically containing a detergent and sometimes a protease.

Protease

An enzyme that digests proteins.

Proteinase K

A broad-spectrum protease used to digest proteins during nucleic acid extraction.

Lysozyme

An enzyme that degrades peptidoglycans in bacterial cell walls, used for lysis of Gram-positive bacteria.

Lysostaphin

An enzyme that cleaves bonds in the cell walls of certain Staphylococcus species, used for lysis.

Lyticase

An enzyme used to digest the cell wall of yeast.

Zymolase

An enzyme preparation used to digest the cell wall of yeast.

Anticoagulant

A substance that prevents blood clotting.

Enzyme inhibitor

A substance that inhibits the activity of an enzyme.

CTAB Extraction Method

A method for extracting DNA using cetyltrimethylammonium bromide, a cationic detergent.

Silica Matrix columns

Columns containing a silica matrix used to selectively bind DNA for purification.

gDNA

Genomic DNA, the total DNA content in a cell or organism.

TE buffer

A buffer solution containing Tris and EDTA, commonly used for storing nucleic acids.

Hydrolysis

The chemical breakdown of a compound due to reaction with water.

Spectrophotometry

A method for measuring the absorbance and transmittance of light through a liquid sample.

UV absorption

The absorption ofultraviolet light by a molecule.

Purine

A class of organic molecules that includes the bases adenine and guanine, found in DNA and RNA.

Pyrimidine

A class of organic molecules that includes the bases cytosine, thymine, and uracil, found in DNA and RNA.

Lambert-Beer law

Law stating that the absorbance of a solution is directly proportional to the concentration of the analyte and the path length of the light beam.

Absorbance

A measure of the capacity of a substance to absorb light of a specified wavelength.

Path length

The distance the light travels through the sample in a spectrophotometer.

Coefficient extinction

A constant that represents the ability of a substance to absorb light at a specific wavelength.

dsDNA

Double-stranded DNA.

RNA

Ribonucleic acid.

ssDNA

Single-stranded DNA.

Nanodrop

A microvolume spectrophotometer used for quantifying DNA, RNA, and protein.

Fluorometry

A method for measuring the fluorescence of a sample.

Fluorescent dye

A dye that emits light of a specific wavelength when excited by light of another wavelength.

EtBr

Ethidium bromide, a fluorescent dye that binds to DNA and is used for visualization.

SYBR Green I

A fluorescent dye that binds to dsDNA.

SYBR Gold

A fluorescent dye that binds to nucleic acids.

PicoGreen

A fluorescent dye that is highly selective for dsDNA.

Fluorometer

An instrument used to measure fluorescence.

Qubit

A type of fluorometer.