promotor studie Flashcards

1
Q

chip

A

= chromatine immunoprecipitatie
= aantonen bep W bind op bep seq in levende cel
5 delen: crosslinking, DNA fragmentatie, immunoprecipitatie, reverse crosslinking, analyse

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2
Q

promotor voorspelling obv seq

A
  • screenen gelijkaardige prom. met orthologen
    > gelijkaardige seq, niet zelfde
    > vergelijken seq, reg elementen in prom.
  • in vivo controle of seq van belang
    > prom in vector prom.loos rapporteergen (promotor probe vector
    > minimale prom = kortste seq prom actief
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3
Q

interpretatie resultaat

A

weinig signaal = seq met rem. RE die TF bind > rem. prom
veel signaal = belangrijkste activiteit in fragment prom
> bio-informatica analyse van kandidaat RE

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4
Q

snelle analyse via

A

RT-qPCR
cDNA micro-array
transcriptomics

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5
Q

controles

A

artificiële situatie ziigdieren
> transient - vals signaal
> RT-qPCR, ChIP (DNA-TF binding) - effect promact
> PC - 2e vector co-transfectie prom-probe vector
eigen rapporteergen
constitutieve prom

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