Lecture 19 Molecular Genetics 2.docx - Lecture 19 Molecular Genetics Oct 29th Labelling single stranded oligonucleotides using polynucleotide

Lecture 19 Molecular Genetics 2.docx - Lecture 19 Molecular...

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Lecture 19: Molecular Genetics Oct 29 th Labelling single stranded oligonucleotides using polynucleotide kinases: - synthesize single stranded oligonucleotide complimentary to a known gene but without phosphates on their 5’ end to locate gene - polynucleotide kinases (PNK) transfer gamma phosphate (radioactively labelled) of ATP to the free hydroxyl of the 5’ of oligonucleotide -can also make them fluorescent instead of radioactive- just have to label them in some way -radiolabelled products need to be rendered single stranded for probe use Southern Blot: analysis of DNA sequence by transferring to a solid-state support (nylon, nitrocellulose) Western blot = for protein detection Process: -cut DNA w/ restriction enzymes so its smaller, make DNA single stranded -separate DNA/RNA by size on gel - transfer to solid state support-sticks to it through covalent bonds- so probes can find specific sequences -hybridize support w/ probes- they find the sequences
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