RNA = onstabiel, boodschap moet niet
lang bestaan.
2’ OH vatbaar voor afbraak door
RNasen (heel stabiel, geen co-factoren
nodig)
3 grote categorieën van RNA die te
maken hebben met DNA-RNA-eiwit
paradigma: mRNA, rRNA, tRNA
(laatste 2 niet coderend)
rRNA = ribosomaal RNA
(verschillende subunits)
tRNA = transfer RNA, fysische link
tussen mRNA en eiwit
Meerdere klassen van niet-coderend RNA ontdekt:
- miRNA
- piRNA = piwi-interacting RNA, interacts with piwi proteins
- siRNA = small interfering RNA, short interfering RNA, silencing RNA bestuderen van
genexpressie
- lncRNA = long non-coding RNA
- circular RNA
, Northern blot: herhaling
Staal RNA extraheren gel blotten op membraan probes
Waarom op membraan en niet in gel?
Probes kunnen er makkelijker bij en ook veel makkelijker om achtergrond ruis weg te doen
Grotendeels vervangen door RNA kwantificeren => kwantitatieve PCR
Termen kennen: Southern (DNA), Northern (RNA), Western (eiwitten) blot
Herhaling: kwantitatieve PCR
Gewone PCR amplificatie met
een eindpunt detectie
kwantitatieve PCR met real-
time na elke cyclus
dPCR, 1 fragment of geen in
putje # positieven tellen
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