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Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
Molecular biology
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Molecular biology

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  • DNA/RNA overview
  • Figure 20.9 Gel electrophoresis
  • Transcript

    • 1. Patricia Linton Manchester Metropolitan University
    • 2. It’s just so amazing – that’s what!
    • 3. It’s not science fiction – it’s science fact! <ul><li>DNA - Encodes all of the genetic information needed for the development and functioning of all cells. </li></ul>Discipline is full of amazing facts
    • 4. TRANSCRIPTION RNA PROCESSING DNA RNA transcript 3  5  RNA polymerase Poly-A Poly-A RNA transcript (pre-mRNA) Intron Exon NUCLEUS Aminoacyl-tRNA synthetase AMINO ACID ACTIVATION Amino acid tRNA CYTOPLASM Poly-A Growing polypeptide 3  Activated amino acid mRNA TRANSLATION Cap Ribosomal subunits Cap 5  E P A A Anticodon Ribosome Codon E The central dogma DNA RNA PROTEIN
    • 5. TRANSCRIPTION TRANSLATION DNA mRNA Ribosome Polypeptide (a) Bacterial cell Nuclear envelope TRANSCRIPTION RNA PROCESSING Pre-mRNA DNA mRNA TRANSLATION Ribosome Polypeptide (b) Eukaryotic cell
    • 6. Uses of molecular biology – endless – revolutionised science <ul><li>Medical </li></ul><ul><li>Production of recombinant proteins for treatment of disease </li></ul><ul><li>Determining the genetic basis of cancer – thereby improving treatment and prognosis </li></ul><ul><li>Diagnosis of disease – fast diagnosis of HIV/TB – infectious diseases </li></ul><ul><li>Agricultural </li></ul><ul><li>pest resistance/ drought resistance/increased productivity </li></ul><ul><li>Industrial </li></ul><ul><li>Manufacture of proteins, strain improvement </li></ul><ul><li>Forensic </li></ul><ul><li>Used to place criminals at scene of crime or rule out suspects – important for conviction </li></ul><ul><li>DNA fingerprinting </li></ul><ul><li>Archaeology </li></ul><ul><li>Analysis of ancient DNA in ceramic food vessels </li></ul><ul><li>Environmental Science </li></ul><ul><li>Identification of species </li></ul><ul><li>Clean-up of oil spills </li></ul>
    • 7. (a) Tobacco plant expressing a firefly gene (b) Pig expressing a jellyfish gene
    • 8. Kary Mullis – Genius or eccentric? ‘ Science grows like a weed every year’ ‘ Each of us have things and thoughts and descriptions of an amazing universe in our possession that kings in the 17th Century would have gone to war to possess’ ‘ Science consistently produces a new crop of miraculous truths and dazzling devices every year’ ‘ I have had an encounter with an extraterrestrial in the form of a fluorescent raccoon’ ??????? You can read his Nobel lecture here: http://nobelprize.org/nobel_prizes/chemistry/laureates/1993/mullis-lecture.html
    • 9. PCR – in the dark ages 8 BORING hours per PCR! 95º C 5 min 35 times 55º C 3 min 72º C 5 min
    • 10. <ul><li>Individual rows of receptacles can be heated – can try lots of reaction temperatures in one ‘run’ </li></ul>
    • 11.  
    • 12. The different steps of PCR <ul><li>Water </li></ul><ul><li>Buffer </li></ul><ul><li>DNA template </li></ul><ul><li>Primers </li></ul><ul><li>Nucleotides </li></ul><ul><li>Mg 2+ ions </li></ul><ul><li>DNA Polymerase </li></ul>PCR recipe
    • 13. The temperature profile of a PCR cycle is controlled by the thermal cycler program which results in a near exponential increase in PCR product accumulation for about the first 30 cycles. The 3 Os!!!!!
    • 14. <ul><li>• Used to separate molecules based on their charge and size </li></ul><ul><li>• Agarose or acrylamide gels can be used to separate DNA fragments </li></ul><ul><li>• DNA is acidic; it migrates from the negative to the positive </li></ul><ul><li>end of the gel </li></ul><ul><li>each fragment’s migration rate is inversely proportional to </li></ul><ul><li>the log of its molecular </li></ul><ul><li>weight </li></ul>
    • 15. http://learn.genetics.utah.edu/content/labs/gel/
    • 16. Mixture of DNA mol- ecules of different sizes Power source Longer molecules Shorter molecules Gel Anode Cathode 1 2 Power source – + + –
    • 17.  
    • 18. DNA Fingerprinting <ul><li>Steps in DNA fingerprinting: </li></ul><ul><ul><li>DNA isolated from tissue sample or PCR’d up </li></ul></ul><ul><ul><li>DNA cut into fragments with enzymes </li></ul></ul><ul><ul><ul><li>DNA with different sequences produce fragments of different sizes </li></ul></ul></ul><ul><ul><ul><li>Can use restriction enzymes to cut either side of VNTRs for genetic typing </li></ul></ul></ul><ul><ul><li>Fragments separated on basis of size and visualized </li></ul></ul><ul><ul><li>Each person’s set of fragments is unique </li></ul></ul>
    • 19.  
    • 20.  
    • 21.  
    • 22. DNA Fingerprinting – identification of the remains of the Russian Royal Family <ul><li>DNA fingerprinting showed that 9 different people were buried in the Ekaterinburg grave. </li></ul><ul><li>Romanovs would be more similar in pattern to each other than to non-relatives. </li></ul><ul><li>All of a child’s bands must be present in one or both of the parents. </li></ul>
    • 23. 7.6 DNA Fingerprinting Adult 1 Adult 2 Adult 3 Adult 4 Adult 5 Adult 6 Child 1 Child 2 Child 3
    • 24. DNA Fingerprinting <ul><li>To see if parents and their children were Romanovs, DNA fingerprints were prepared for relatives of tsar and tsarina. </li></ul><ul><li>Adult male skeleton (related to the children) was related to George, the tsar’s brother. </li></ul><ul><li>Adult female skeleton (related to the children) was related to Prince Philip, the tsarina’s grand-nephew. </li></ul><ul><li>Conclusion : the grave contained the tsar, tsarina, three of their children, and four servants. </li></ul>
    • 25. Making your own equipment
    • 26.  

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