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In Vitro Mutagenesis
technique used to induce certain mutation into a cloned DNA fragment
done outside of living organism
done to see the impact of protein being made/not made
ex. GMOs, medical research, study of human disease
In Situ Hybridization
allows scientists to visualize the location of a specific DNA or RNA sequence within a tissue sample
highly dependent on base pairing
CISH
chromogenic ISH
dependent on antibodies and enzymes
done with light microscope, so cheaper
FISH
fluorescence ISH
higher sensitivity
more expensive
used to test for colon cancer, breast cancer, HPV, other viruses
Gel Electrophoresis
used to separate DNA and proteins based on size and charge
requires restriction enzymes, gels, power supply and staining
used for forensics and paternity testing
ELISA
enzyme linked immunosorbent assay
used to detect and quantify proteins, antibodies, and antigens using enzyme linked reactions
used to test testosterone levels in athletes, food for allergens, and antibody testing
Next Generation Sequencing
used for DNA and RNA sequencing
can sequence multiple strands at one time
main application is in microbiology
Forensic Profiling
using DNA sample for identification
highly based on short tandum repeats
ex. maternity/paternity testing, body identification
Stem Cells
undifferentiated/unspecialized cells that have the potential to become any type of cells
Totipotent Cells
can develop into any type of cell
Pluripotent Cells
can only develop into some types of cells
DNA Microarray Assay
procedure that uses DNA or RNA in fragments to hybridize two complementary strands and attach florescent markers on a chip
uses include identifying specific types of cancer and to determine which genes are on vs off
CRISPR-Cas 9
cut and past for DNA
cancer, cystic fibrosis, sickle cell disease, agriculture
GMOs
genetically modified organisms
controversial due to food safety and environmental impacts
DNA cloning
possible for gene, cell, or total organism
Dolly the Sheep
organ donation, create biodiversity, agriculture and animals
PCR
polymerase chain reaction
denaturation, annealing, extension
makes more copies of DNA
RT PCR —> copies of complementary DNA made from RNA