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Efteruddannelse i Bioteknologi

Efteruddannelse i Bioteknologi. Kursus A PCR af gDNA. Kursus indhold. PCR af plante gDNA. Baseret på kittet: GAPDH/PCR Module PCR: http://www3.bio-rad.com/LifeScience/jobs/2004/04-0522/04-0522_PV92_PCR.html. PCR. PCR. PCR. PCR. GAPDH Glyceraldehyde 3-phosphate dehydrogenase.

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Efteruddannelse i Bioteknologi

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  1. Efteruddannelse i Bioteknologi Kursus A PCR af gDNA

  2. Kursus indhold

  3. PCR af plante gDNA Baseret på kittet: GAPDH/PCR Module PCR: http://www3.bio-rad.com/LifeScience/jobs/2004/04-0522/04-0522_PV92_PCR.html

  4. PCR

  5. PCR

  6. PCR

  7. PCR

  8. GAPDHGlyceraldehyde 3-phosphate dehydrogenase

  9. Flere gener for ”samme” enzym

  10. GAPDH GAPDH katalyserer reaktionen: Glyceraldehyde-3-phosphate + NAD+ + Pi  1,3-bisphosphoglycerate + NADH + H+

  11. I Arabidopsis

  12. GAPDH tetramer

  13. Primer design

  14. Primer design • Længde • Smelte temperatur Tm • Tm = 2°C· (A+T) + 4°C· (G+C) • Annealing temperatur • Primer hairpins • Primer dimers • GC-clamp

  15. PCR kloning

  16. Degenerede primere • Kendte sekvenser • Nye homologe gener med næsten ens sekvenser • Primere som næsten passer • Blandet population af primere

  17. Primer design

  18. Primer design

  19. Primer design

  20. Exonukleasebehandling • Nedbrydning af primere fra 1. runde PCR • Enkeltstrengs nuklease • Dobbeltstrenget PCR-fragment overlever

  21. Nestede primere

  22. Nestede primere

  23. Gennemgang af forsøgsvejledning Se i mappen

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