Ch.20-biotechnologyrevision.ppt-byrdistheword.ppt

* * * * * * * * * * * PCR is an incredibly versatile technique: An important use of PCR now is to “pull out” a piece of DNA sequence, like a gene, from a larger collection of DNA, like the whole cellular genome. You don’t have to go through the process of restriction digest anymore to cut the gene out of the cellular DNA. You can just define the gene with “flanking” primers and get a lot of copies in 40 minutes through PCR. Note: You can also add in a restriction site to the copies of the gene (if one doesn’t exist) by adding them at the end of the original primers. * * Taq = Thermus aquaticus (an Archaebactera) Highly thermostable – withstands temperatures up to 95°C for more than 40min. BTW, Taq is patented by Roche and is very expensive. Its usually the largest consumable expense in a genomics lab. I’ve heard stories of blackmarket Taq clones, so scientists could grow up their own bacteria to produce Taq in the lab. It’s like pirated software -- pirated genes! * In 1985, Kary Mullis invented a process he called PCR, which solved a core problem in genetics: How to make copies of a strand of DNA you are interested in. The existing methods were slow, expensive imprecise. PCR turns the job over to the very biomolecules that nature uses for copying DNA: two primers that flag the beginning end of the DNA stretch to be copied; DNA polymerase that walks along the segment of DNA, reading its code assembling a copy; and a pile of DNA building blocks that the polymerase needs to make that copy. As he wrote later in Scientific American: Beginning with a single molecule of the genetic material DNA, the PCR can generate 100 billion similar molecules in an afternoon. The reaction is easy to execute. It requires no more than a test tube, a few simple reagents and a source of heat. The DNA sample that one wishes to copy can be pure, or it can be a minute part of an extremely complex mixture of biological materials. The DNA may come from a hospital tissue specimen, from a si