Biotechnology (Chapter 20)

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