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In Situ Hybridization



In situ hybridization helps detecting and localizing RNA messengers on paraffin-embedded sections, using complementary RNA labeled probes.




The probes hybridize with sequence of interest, so they can be visualized in optic microscopy after enzymatic revealing. This method completes the immunochemistry technique which highlights a specific protein.

To improve its knowledge in tissular characterization, Novotec has undertaken the development of in situ hybridization since 2010. From tissue to cell, from protein to messengers, Novotec is able to comprehend the entirety of cellular mechanisms.

Skin - In situ hybridization with col3a1 RNA probeSkin - <em>In situ</em> hybridization with col1a1 RNA probeCutaneous substitute - <em>In situ</em> hybridization with vegf-a RNA probe

Novotec offers a personalized service at each step :

  • Sequence design : GenBank Accession Number, Gene ID…

  • Probe making, labeling and purification : RNA probe (sense et antisense); digoxigenin labeling. Possibility to use Novotec’s validated probes (col1a1, col3a1, vegf-a…)

  • In situ hybridization : optimization of the various hybridization parameters contigent on the tissue and on the probe.

chromatogramm (Dna baser v2)






References

  • «In situ Hybridization to evaluate the wound healing process of a bilayered living cell-based construct (Apligraf®) in a nude mice model - 2010 june, TERMIS-EU, Galway, Ireland.»
    Rousseau C. and Pagnon-Minot A., Guerret S., Hartmann D.J, Ronfard V.
  • «Short term integration on human bilayered living skin substitute (Apligraf®) grafted onto nude mice - 19th Annual Meeting of the Wound Healing Society with SAWC (Symposium on Advanced Wound Care) 2009 April 26-29, Gaylord Texan Resort and Convention Center, Dallas, Texas.»
    Rousseau C., Thillou F., Pagnon-Minot A., Guerret S., Hartmann D.J, Ronfard V.
  • «Characterization of spatial and temporal expression of pattern of Col15a1b during zebrafish development. Gene Expression Patterns 2011, 11, 129-134.»
    Bretaud S., Pagnon-Minot A.,Guillon E., Ruggiero F., Le Guellec D.
  • «Collagen XV, a novel factor in zebrafish notochord differentiation and muscle development. Dev Biol 2008, 316, 21-35.»
    Pagnon-Minot A., Malbouyres M., Haftek-Terreau Z., Kim H.R., Sasaki T., Thisse C., Thisse B., Ingham P.W., Ruggiero F., Le Guellec D.