Gene isolation and amplification
Gene isolation and amplification Biology 215
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This 2 page Class Notes was uploaded by Skylar on Monday May 16, 2016. The Class Notes belongs to Biology 215 at Northwestern University taught by Galio in Spring 2016. Since its upload, it has received 8 views. For similar materials see Genetic and Molecular Biology in Biology at Northwestern University.
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Date Created: 05/16/16
Gene Isolation and Manipulation Overview -First step of studying gene function- isolate its DNA and reproduce it in large quantites. -One way is to cut the genome with “molecular scissors” aka restriction endonucleases and isolate the fragments containing the desired gene. -DNA Technology: the techniques used for obtaining, amplifying, and manipulating specific DNA fragments. -Genetic Engineering: application of DNA technology to specific biological, medical or agricultural problems. -Genomics: global analysis of the nucleic acids present in a nucleus, a cell, an organism or a group of related species. Isolating and Amplifying Specific DNA fragments -Amplification: replicate the DNA the desired segment into large quantities. -In vivo: -Researcher uses Donor DNA: sample of DNA molecules containing the gene of interest. Usually an entire genome. -Vectors: plasmids or bacterial viruses that can “carry” the inserted DNA fragments. - restriction endonucleases cut long chromosome size DNA molecules into hundreds or thousands of fragments. -the fragments are inserted into a cut vector that can fit the fragment- this forms recombinant DNA -The recombinant DNA is inserted into bacterial cells- usually only one is taken up by the cell. -the DNA is amplified and during cell division which results in a clone of identical cells each containing the recombinant DNA- this is called DNA cloning -Many fragments of the DNA inserted into the vector meaning the recombinant DNA has almost as much as the entire genome of the original DNA. Need to pinpoint the gene we want. -In vitro: -aka polymerase chain reaction (PCR) -a specific gene or DNA region of interest is isolated and amplified by DNA polymerase. -PCR finds the target DNA due to the help of primers. -PCR results in large quantities of the target DNA -DNA tech depends on: -the ability of specific proteins to recognize and bind to specific bases sequences in DNA -ability of complementary single stranded DNA or RNA sequences to anneal to form double stranded molecules -Example of the use of recombinant DNA -Diabetes- cow have a gene that makes insulin protein easily. Researchers isolated that gene and were able to make recombinant human insulin to help those with diabetes.
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