{"id":91,"date":"2020-08-20T16:43:50","date_gmt":"2020-08-20T20:43:50","guid":{"rendered":"https:\/\/pressbooks.bccampus.ca\/kathleef\/?post_type=chapter&p=91"},"modified":"2025-06-16T16:20:40","modified_gmt":"2025-06-16T20:20:40","slug":"chapter-16-applications-of-pcr","status":"publish","type":"chapter","link":"https:\/\/pressbooks.bccampus.ca\/kathleef\/chapter\/chapter-16-applications-of-pcr\/","title":{"raw":"Further Applications of PCR","rendered":"Further Applications of PCR"},"content":{"raw":"

Introduction<\/h1>\r\nIn this section we'll look at some applications of PCR, beyond those already covered in the alternative cloning approaches and traditional cloning chapters. You will already appreciate that we can add sequences to PCR primers so that we can amplify products that can be cut with certain enzymes or generate overhangs for ligase independent cloning.\r\n\r\nBut we can use PCR for other purposes as well.\r\n\r\nIn terms of making transgenic organisms, we can use PCR to introduce mutations into DNA sequences in order to study the function of the gene.\u00a0 We can also use PCR to fuse two sequences together- as always, primer design is key here and the primers include sequence overlaps between the two pieces to be combined (you have seen the same concept in the SLIC cloning chapter).\r\n\r\nOnce the transgenic organism is produced, we can also use PCR as a way to confirm the presence of the transgene in the organism, determine the exact location of the transgene, and test individuals to find out if they are homozygous or heterozygous for the inserted DNA.\r\n\r\nThis recording covers a few basics about add<\/span>ing nucleotides to primers for cloning purposes. An example is adding restriction sites to primers so that an insert can be cloned directionally. It is mainly review. <\/span>Click here for the powerpoint slides<\/a>.<\/span>\r\n\r\n[embed]https:\/\/www.youtube.com\/watch?v=oBNHhz5z4eE&feature=youtu.be&hd=1[\/embed]\r\n
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Contents<\/h2>\r\n

Learning Outcomes<\/a>\r\nA. Site-directed mutagenesis<\/a><\/p>\r\n

A-1. Types of changes that can be introduced<\/a>\r\nA-2. Using the genomic DNA as template<\/a>\r\nA-3. Using the cloned gene in a plasmid as a template<\/a><\/p>\r\n

B. Fusion PCR<\/a>\r\nC. PCR on transgenic organisms<\/a><\/p>\r\n

C-1. Confirmation of the presence of the foreign DNA<\/a>\r\nC-2. Determining the position of the transgene<\/a>\r\nC-3. Checking for homozygosity or heterozygosity<\/a><\/p>\r\n \r\n\r\nList of lecture videos (excluding supplemental videos)<\/a>\r\n\r\n<\/div>\r\n\r\n

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<\/a>Learning Outcomes<\/h2>\r\n<\/header>\r\n
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