Biotechnology Flashcards
(10 cards)
step 1: isolation of DNA (restriction enzymes)
A restriction enzyme (endonuclease) is a DNA-cutting enzyme that recognises a specific target sequence and cuts DNA into two pieces at or near that site (recognition site)
step 2: insertion of DNA (plasmid vector)
Use the same restriction enzyme that you used to remove the DNA fragment in step 1 to make a single cut in a plasmid. This is going to be the plasmid vector that is inserted back into a bacterium
step 3: joining of DNA (DNA ligase)
DNA ligase seals gaps in DNA backbone. Therefore, when it’s used in recombinant DNA, it will link the plasmid and the DNA fragment to make a recombinant plasmid containing the gene.
DNA ligase links the phosphate group of one DNA strand to the hydroxyl group of the other DNA strand to form a single sugar-phosphate backbone/combined piece of DNA
step 4: amplification of recombinant DNA (bacterial transformation)
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step 5: produce your protein
When a bacterial colony with the right plasmid has been found, then it can be grown in a culture
The bacteria start to transcribe the gene and will produce the coded for functional protein.
The bacteria are then split open to release the product and the end product is then purified
DNA sequencing - mapping genomes
DNA sequencing is the process of determining the nucleic acid sequence in DNA.
Applications of DNA sequencing: mapping species genomes
Complete sequences of genomes allow us to compare the DNA of closely related species in order to establish and analyse their genetic differences, a field known as comparative genomics.
DNA profiling - identifying info
DNA profiling is the process of determining an individual’s DNA characteristics.
Applications of DNA profiling: to identify unique genetic information
DNA profiling is a forensic technique in criminal investigations, comparing criminal suspects’ profiles to DNA evidence so as to assess the likelihood of their involvement in the crime. It is also used in parentage testing, to establish immigration eligibility, and in genealogical and medical research.
Polymerase chain reaction (PCR)
enables us to extract a tiny quantity of DNA from a single hair or drop of blood at the scene of a crime and increase the amount of it a million times or more so that it can be analysed.
Allows us to analyse the DNA from a bacterial or viral infection in order to diagnose an illness.
Makes it possible to mass-produce DNA from fossil remains, extinct for millions of years.
gel electrophoresis
To separate DNA fragments, based on size.
Can be used to determine paternity or criminals: DNA profiling.
DNA molecules are negatively charged, so if you apply an electrical current to a medium they are present in, they will move towards the positive electrode
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