DNA manipulation

Biotechnology

use of living things and biological systems for human benefit

Genetic engineering

Manipulation of genetic material in an organisms through modern science

GMO

Genetically modified organism resulting from biotechnology techniques

transgenic organism

GMO that has been modified by having foreign DNA from another species inserted

Knock out organisms

cutting out genes or gene segments to prevent expression or functioning of gene product, usually through a frameshift mutation.

Knock in organisms

inserting genes in a specific gene locus to be controlled by a specific promotor

reporter gene

gene inserted to be expressed under the same promoter as gene of interest, provides info about location and timing of a gene of interest

restriction endonucleases/ enzymes

used to cut DNA sugar-phosphate backbone, recognises specific restriction site of 4-8 base pairs. Originally isolated from bacteria where they cut viral DNA

sticky ends

Uneven cut of DNA leaving a single stranded tail that will only join with another complementary sticky end

blunt ends

even cut of DNA that can join with any other blunt end piece- non-specific

DNA ligase

joins two pieces of DNA together using phosphodiester bonds. Found naturally in cells, can join fragments from different organisms if sticky/blunt ends are complementary.

recombinant DNA

DNA that has been recombined from different sources using DNA ligase. Segments must be cut with same endonuclease to produce complementary sticky ends

CRISPR-Cas9 in bacteria

Viruses infect bacteria by inserting its DNA, bacteria collects fragments of viral DNA and inserts

crRNA

cas9 protein

PAM sequence

sgRNA

CRISPR-cas9 biotechnology

editing disease genes with CRISPR

CRISPR in agriculture

CRISPR mouse strains

CRISPR ethics

PCR

PCR requirements

primers

Denaturation in PCR

Annealing in PCR

Extension in PCR

Gel electropheresis

Molecular size markers

DNA profiling

Short tandem repeats

DNA profiling ethics

recombinant plasmids

steps to make recombinant plasmids

vectors

gene cloning

antibiotic resistance genes

promoter

gene cloning applications

gene cloning ethics

Insulin production recombinant plasmids

gel electrophoresis and insulin production

insulin detection with antibiotic

insulin detection with beta-galactosidase