{"id":5262,"date":"2020-03-03T12:00:57","date_gmt":"2020-03-03T20:00:57","guid":{"rendered":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/?p=5262"},"modified":"2020-03-05T21:26:23","modified_gmt":"2020-03-06T05:26:23","slug":"what-is-pcr","status":"publish","type":"post","link":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/2019ncov\/what-is-pcr\/","title":{"rendered":"What is PCR?"},"content":{"rendered":"\n

PCR stands for “polymerase chain reaction”. It was discovered in the 1983 by Kary Mullis. PCR is a technique that is used to create a whole bunch of copies (amplify) of a section of DNA or a gene for studying. <\/p>\n\n\n\n

Download the high-quality<\/a> <\/a>poster here.<\/a><\/p>\n\n\n\n

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PCR “Recipe”<\/h3>\n\n\n\n

Ingredients: <\/p>\n\n\n\n

  • DNA template <\/strong>(this is what you want to copy).<\/li>
  • Primers <\/strong>that start the PCR reaction (on either side of the section of DNA you want copied). These primers are specific for the piece of DNA you want to copy.<\/li>
  • You need the building blocks<\/strong> which are the nucleotide bases (Adenine, Cytosine, Guanine, Thymine<\/strong>) so that you can make more copies of the DNA.<\/li>
  • Thermostable DNA polymerase, Taq polymerase<\/strong> (T<\/strong>hermus aq<\/strong>uaticus<\/em>) enzyme. Taq polymerase comes from the bacteria Thermus aquaticus<\/em> or T. aquaticus<\/em>. Thermostability is important because the PCR process involves repeated cycles (20-40 cycles) of heating and cooling.<\/li>
  • Buffer <\/strong>to help stabilize\/maintain conditions for the enzyme to work to build copies that you want.<\/li><\/ul>\n\n\n\n

    Thermocycling (repeat steps 1-3, 20-40 times)<\/h3>\n\n\n\n

    Step 1. Denaturing. Heat to 94-96 degrees C. The heat will help break the hydrogen bonds thus separating the two strands of DNA. This takes 15-30 seconds.<\/p>\n\n\n\n

    Step 2. Annealing. Cool to 50-65 degrees C. This allows the primers to bind to the individual strands of DNA via hydrogen bonding. This takes 10-30 seconds.<\/p>\n\n\n\n

    Step 3. Extension. Heat to 72 degrees C. The T. aquaticus<\/em> bacteria is a heat-lover which allows it to withstand and function at higher temperatures. Taq<\/em> polymerase attaches to primers and then adds bases, one-at-a-time, from the 5′ to 3′. The approximate rate is 1000 DNA bases (1Kb, kilobase) per 1 minute.<\/p>\n\n\n\n

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