* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download DNA Cloning - MrMsciences
Genome evolution wikipedia , lookup
DNA barcoding wikipedia , lookup
Epigenetics wikipedia , lookup
Epigenetics in learning and memory wikipedia , lookup
Human genome wikipedia , lookup
Gene therapy wikipedia , lookup
Comparative genomic hybridization wikipedia , lookup
Mitochondrial DNA wikipedia , lookup
Metagenomics wikipedia , lookup
DNA profiling wikipedia , lookup
DNA polymerase wikipedia , lookup
Zinc finger nuclease wikipedia , lookup
SNP genotyping wikipedia , lookup
Primary transcript wikipedia , lookup
Cancer epigenetics wikipedia , lookup
Nutriepigenomics wikipedia , lookup
Genetic engineering wikipedia , lookup
Bisulfite sequencing wikipedia , lookup
Point mutation wikipedia , lookup
DNA damage theory of aging wikipedia , lookup
United Kingdom National DNA Database wikipedia , lookup
Genealogical DNA test wikipedia , lookup
Nucleic acid analogue wikipedia , lookup
Microsatellite wikipedia , lookup
Gel electrophoresis of nucleic acids wikipedia , lookup
No-SCAR (Scarless Cas9 Assisted Recombineering) Genome Editing wikipedia , lookup
Cell-free fetal DNA wikipedia , lookup
Non-coding DNA wikipedia , lookup
Genomic library wikipedia , lookup
DNA vaccination wikipedia , lookup
Nucleic acid double helix wikipedia , lookup
DNA supercoil wikipedia , lookup
Site-specific recombinase technology wikipedia , lookup
Epigenomics wikipedia , lookup
Genome editing wikipedia , lookup
Extrachromosomal DNA wikipedia , lookup
Microevolution wikipedia , lookup
Designer baby wikipedia , lookup
Cre-Lox recombination wikipedia , lookup
Deoxyribozyme wikipedia , lookup
Therapeutic gene modulation wikipedia , lookup
Vectors in gene therapy wikipedia , lookup
Molecular cloning wikipedia , lookup
Helitron (biology) wikipedia , lookup
DNA Cloning Cloning • a line of genetically identical cells or individuals derived from a single ancestor • produces many copies of a piece of DNA • uses a little fraction as gene of interest • cultivates a large amount for studying functions Functions • Biological • determine the DNA sequence • ultimate information • regulation • Pharmaceutical • manufacture medicines • Clinical How it Works • Combine gene of interest and bacterial plasmid • Recombinant DNA • DNA from two or more different sources that have been joined together to form a single molecule • Amplification and identification Zoom In!!! • Insertion, the most critical step • made possible by restriction endonucleases(aka. restriction enzymes) • recognize specific DNA sequences called restriction sites • cut the DNA at specific locations • Restriction fragments Restriction Enzyme • defend against viral attack by breaking down the DNA molecules of infecting viruses • cleave the sugar-phosphate backbones of DNA to produce sticky ends • short single-stranded regions • form hydrogen bonds with complementary sticky ends on any other DNA molecules cut with the same enzyme • kind of like glue when you fix something Cloning Vectors • DNA molecules into which a DNA fragment can be inserted to form a recombinant DNA molecule for the purpose of cloning • plasmid cloning vectors • contains 2 unique genes: ampR gene and lacZ+ gene The Two Genes • ampR gene • encodes an enzyme that breaks down the antibiotic ampicillin • lacZ+ gene • encodes ßgalactosidase (hydrolyzes the sugar lactorse and synthetic substrates)