WET LAB DNA Barcoding: From Samples to Sequences - Northwest ...

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PCR beads: Lyophilized or “freeze-dried” PCR ingredients.
PCR product: The DNA copied or “amplified” during PCR.
PCR purification: Extracting or purifying the PCR product (DNA fragment) away from the remaining PCR components.
Pellet: After centrifugation, the pellet is the material found at the bottom of the tube.
Phosphodiester bond: A phosphodiester bond is a strong covalent bond between a phosphate group and two five-carbon
ring carbohydrates. DNA polymerase catalyzes the formation of phosphodiester bonds between the 3’ hydroxyl group
on the deoxyribose at the end of a DNA strand and the phosphate group attached to the 5’ carbon of the deoxyribose on
the new DNA nucleotide.
Polymerase Chain Reaction (PCR): A scientific technique in molecular biology used to amplify (i.e., copy) a single
or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a
particular DNA sequence.
Power supply: Used to create the electrical field to which the DNA will be subjected during agarose gel electrophoresis.
Primer pools: Collections or mixtures of primers, usually used in PCR.
Primers: Small pieces of DNA used to start or “prime” DNA synthesis, as in DNA replication and PCR.
Proteinase K: Type of enzyme that breaks down proteins, including nucleases.
Reagents: Ingredients or components used in an experiment.
Sanger sequencing: A method of DNA sequencing that uses deoxyribonucleaotides (dNTPs) and
dideoxyribonucleotide triphosphates (ddNTPs). The ddNTPs end DNA synthesis at random positions, and the resulting
fragments can be analyzed to determine the exact DNA sequence. For more on Sanger sequencing, see also DNA
sequencing.
Spin column: A small column that fits inside of a microfuge tube and contains a material (such as silica) that binds to
nucleic acids such as DNA. The spin column is used in conjunction with centrifugation (or vacuum pressure) to purify DNA.
Supernatant: After centrifugation, the supernatant is the liquid found above the pellet.
Taq DNA polymerase: A type of heat-stable DNA polymerase, purified from the thermophilic (“heat-loving”) bacteria
Thermus aquaticus.
Template: See DNA template.
Thermocycler: Type of machine used to automate polymerase chain reaction that cycles through all of the temperatures
required to complete the PCR.
Total DNA: All of the DNA found in a cell, including the DNA in the nucleus and in the mitochondria.
Wells: Holes in a gel into which samples are placed or “loaded.”
3’ hydroxyl group: During DNA synthesis, DNA polymerase catalyzes the formation of a phosphodiester bond
between the 3’ hydroxyl group on the deoxyribose at the end of a DNA strand and the phosphate group attached to the 5’
carbon of the deoxyribose on the new DNA nucleotide. See also DNA sequencing.
Wet Lab – DNA Barcoding: From Samples to Sequences
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©Northwest Association for Biomedical Research—Updated October 2012