Lab 4 Flashcards
…: the science of classifying living things according to shared features
taxonomy
carl Linnaeus formalized biological classification with his system of … that assigns each organism a genus and species name
binomial nomenclature
identifying organisms has grown in importance as we monitor the biological effects of … and attempt to preserve … in the face of accelerating habitat destruction
global climate change; species diversity
scientists agree that the yearly rate of extinction has increased from about … species per million to …-… species per million
1; 100-1000
… allow non-experts to objectively identify species
DNA barcodes
a DNA barcode is a … that identifies each living thing. short DNA barcodes, about 700 nucleotides in length, can be quickly processed from thousands of specimens and unambiguously analyzed by computer programs
unique pattern of DNA sequence
the 10-year …, completed in 2010, provided the first comprehensive list of more than 190,000 marine species and identified 6000 potentially new species
census of marine life
barcoding relies on …, highly … regions of the genome
short; variable
a region of the chloroplast gene … is used for barcoding plants
rubisco large subunit
a region of the mitochondrial gene … is used for barcoding animals
cytochrome c oxidase
the genes used for barcoding are involved in the key reactions of life: storing … in … and releasing it to form ..
energy; carbs; ATP
first, a sample of … is collected, preserving the specimen whenever possible and noting its … and …
tissue; geographical location; local environment
DNA is extracted from a small sample and the barcode portion of the rbcL gene is amplified by … the amplified sequence is submitted for sequencing in one or both directions
PCR
the sequencing results are then used to search a …
DNA database
put sample into ziplock with a small amount of … to keep it fresh
salt
(DNA extraction) …: the DNA is enclosed within the cell and nuclear membranes so the first step is to get the DNA out. Cells will be lyced. The sample will be macerated and then we will use a … to break up the lipids The enzyme … must be added to denature hydrolytic proteins and keep the DNA intact.
cell lysis; soapy lysis buffer (AP1); RNase A
(DNA extraction) …: once you have removed the hydrolytic enzymes you can begin the DNA … process. In essence you will put the cellular components into a spin column to remove the DNA. The DNA sticks to a … across the column, the proteins and other cytoplasmic components pass through. this is accomplished by adding … and … to your solution
elimination of cellular debris; purification; membrane; ethanol; protein binding buffer
(DNA extraction) elimination of cellular debris contd:
upon centrifugation, this protein precipitating solution will pass through the membrane that will attract the DNA and allow debris to pass through. two … steps follow this with the addition of buffers AW1 and AW2 to clean the DNA from remaining protein debris
wash
(DNA extraction) 3. …: you will now remove the DNA from the matrix by adding an elution (…) buffer, buffer AE
DNA elution; removal
