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3C-RT-qPCR?



  1. #1
    MaliciaR

    3C-RT-qPCR?


    ------

    Bonjour,

    J'étais à un séminaire l'autre jour et le garçon a parlé d'une technique appelée 3C couplée (pour l'analyse des résultats) à une RealTime-qPCR. Quelqu'un saurait-il de quoi il retourne?

    Merci

    Cordialement,

    -----
    An expert is one who knows more and more about less and less.

  2. #2
    MaliciaR

    Re : 3C-RT-qPCR?

    J'ai trouvé : 3C = Chromosome Conformation Capture
    Voici l'article du protocole initial (http://www.ncbi.nlm.nih.gov/pubmed/1...ubmed_RVDocSum) :


    Quantitative analysis of chromosome conformation capture assays (3C-qPCR).

    Hagège H, Klous P, Braem C, Splinter E, Dekker J, Cathala G, de Laat W, Forné T.

    Chromosome conformation capture (3C) technology is a pioneering methodology that allows in vivo genomic organization to be explored at a scale encompassing a few tens to a few hundred kilobase-pairs. Understanding the folding of the genome at this scale is particularly important in mammals where dispersed regulatory elements, like enhancers or insulators, are involved in gene regulation. 3C technology involves formaldehyde fixation of cells, followed by a polymerase chain reaction (PCR)-based analysis of the frequency with which pairs of selected DNA fragments are crosslinked in the population of cells. Accurate measurements of crosslinking frequencies require the best quantification techniques. We recently adapted the real-time TaqMan PCR technology to the analysis of 3C assays, resulting in a method that more accurately determines crosslinking frequencies than current semiquantitative 3C strategies that rely on measuring the intensity of ethidium bromide-stained PCR products separated by gel electrophoresis. Here, we provide a detailed protocol for this method, which we have named 3C-qPCR. Once preliminary controls and optimizations have been performed, the whole procedure (3C assays and quantitative analyses) can be completed in 7-9 days.


    Et j'ai vu qu'il y a aussi 5C, mais bon, les 3 me suffisent pour l'instant


    Cordialement,
    An expert is one who knows more and more about less and less.

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