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Laser capture microdissection of intestinal tissue from sea bass larvae using an optimized RNA integrity assay and validated reference genes. <i>NPG Scientific Reports 6</i>: 21092. <a href=\"https://dx.doi.org/10.1038/srep21092\" target=\"_blank\">https://dx.doi.org/10.1038/srep21092</a>","BEntID":248704,"PublicFlag":1,"CheckedFlag":1,"wosflag":1,"vabbflag":1,"RefStringPartII":". <i>NPG Scientific Reports 6</i>: 21092. <a href=\"https://dx.doi.org/10.1038/srep21092\" target=\"_blank\">https://dx.doi.org/10.1038/srep21092</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Schaeck, M.; De Spiegelaere, W.; De Craene, J.; Van Den Broeck, W.; De Spiegeleer, B.; Burvenich, C.; Haesebrouck, F.; Decostere, A.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Schaeck, M. <i>et al.</i>","Englishabstract":"The increasing demand for a sustainable larviculture has promoted research regarding environmental parameters, diseases and nutrition, intersecting at the mucosal surface of the gastrointestinal tract of fish larvae. The combination of laser capture microdissection (LCM) and gene expression experiments allows cell specific expression profiling. This study aimed at optimizing an LCM protocol for intestinal tissue of sea bass larvae. Furthermore, a 3'/5' integrity assay was developed for LCM samples of fish tissue, comprising low RNA concentrations. Furthermore, reliable reference genes for performing qPCR in larval sea bass gene expression studies were identified, as data normalization is critical in gene expression experiments using RT-qPCR. We demonstrate that a careful optimization of the LCM procedure allows recovery of high quality mRNA from defined cell populations in complex intestinal tissues. According to the geNorm and Normfinder algorithms, <i>ef1a, rpl13a, rps18</i> and <i>faua</i> were the most stable genes to be implemented as reference genes for an appropriate normalization of intestinal tissue from sea bass across a range of experimental settings. 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