Biotechnology

• Biotechnology


  The use of an organism or other biological system to make a product or process
• DNA technology


  Approaches to the sequencing and manipulation of DNA
• Vector


  Piece of carrier DNA used in gene therapy to get it into cells so it can be expressed
• Clone


  To make an exact genetic copy of something. Can be to an entire organism, or just to tiny bits of DNA and individual genes
• DNA cloning


  The process of making multiple identical copies of a particular piece of DNA by recombining it into a plasmid, which is put into bacteria and then reproduced when they do. Good for doing things like farming insulin or other specific proteins
• Recombinant DNA


  DNA assembled from multiple sources
• Restriction enzyme


  DNA cutting enzyme that recognizes a specific target and cuts near it, often leaving single-stranded overhangs. If overhangs match, the string can be reattached using ligase so now you have modified DNA
• Transformation


  When bacteria are given a shock that makes them more likely to take up a plasmid (which was modified during the cutting and pasting of DNA phase of cloning)
• Plasmids typically contain an antibiotic resistance gene, so the bacteria exposed to the plasmids are exposed to antibiotics they give resistance to. It will kill all the ones without the plasmid and its resistance, so only the ones with the plasmid, its resistance, and your cloned gene will survive and get turned into colonies
• Once bacteria containing the cloned gene are in a big enough colony, they are split open to free the desired proteins, and then purified so that it can be experimented on or given to patients
• Taq polymerase


  DNA polymerase typically used in PCR because of its heat stability and the fact that it works best at 70 degrees C, because PCR typically has to expose DNA to high temps to denature it
• Main PCR steps:

  • Denaturing
  • Annealing
  • Extension
• Denaturing


  1st step of PCR, when you heat up the DNA so its strands separate, leaving you with single-stranded templates
• Annealing


  2nd step of PCR, when you cool the reaction so the primers can find their complimentary sequences on denatured single-strand DNA
• Extension


  3rd step of PCR, when temperatures are raised so taq polymerase extends the primers, synthesizing new cloned strands of DNA
• Gel electrophoresis


  Technique used to separate DNA fragments according to their size
• Gel electrophoresis


  Technique where particles move differently through a gel depending on their size and charge when a current is ran through it. Because all DNA has the same charge, all pieces of it move solely according to their size
• Agarose


  Polysaccharide material commonly used in gel electrophoresis
• DNA ladder


  Assortment of DNA at known lengths that is used as a standard for the target fragments to be measured against in gel electrophoresis
• Band


  Well-defined line of DNA on the gel in gel electrophoresis
• DNA sequencing


  Determining the sequence of nucleotide bases in a piece of DNA
• Sanger sequencing


  When a bunch of DNA clones are put into a PCR reaction but with a very small amount of chain-ending dyed nucleotides. Through probability, each nucleotide should have a chain end on it at least once. Then, they are put through gel electrophoresis to determine which strands end on what in what order to sequence them
• Chromatogram


  Device used to show peaks in fluorescent activity to determine which nucleotides go where in sanger sequencing
• Next generation sequencing


  The collective term for newer DNA sequencing technologies