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  • General Question: Is it possible to generate mRNA from genomic DNA (in vitro)?

    Hello,

    If I have isolated genomic DNA, is it possible to induce transcription via some commercially available kit? If this is possible, is there a way to get every mRNA transcript that could be found potentially be found in vivo ?

    Thanks!

  • #2
    Assuming this is an organism in which substantial post-translational processing occurs, and you want to get something like the final, spliced mRNAs, I would have to guess "no". If you just want to convert the whole genome from DNA into RNA, that should be possible.

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    • #3
      No. Isolated genomic DNA is denatured. Only undenatured genomic DNA in situ can support all possible in vivo transcripts. You can look to isolated protein and amino acid sequences as ex vivo recordings of possible transcript variants.

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      • #4
        Even theoretically it is not possible to get the complete functional transcriptome from isolated genomic DNA; main reasons being: complex organization of the eukayotic genes (introns/exons), and a general absence of universal start signals for transcription of genes across the whole genome.

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