gene expression analysis: rt-pcr

ZF BioLabs analyzes the expression of a specific zebrafish gene by RT-PCR, during the first five days of embryonic development. For this purpose, the sequence of the zebrafish homologous gene has to be identified from published sequences in zebrafish databases. If the gene of interest has not been previously described in zebrafish, its sequence should be searched among the different orthologous published. In both cases and with the supplied sequence, ZF BioLabs will design specific oligonucleotides to amplify the target gene. Total zebrafish isolated RNA, from 1, 2, 3, 4 and 5 days post fertilization embryos, will be used to carry out the RT-PCR. As an internal control, primers for a standard housekeeping gene (β-actin) will be included.

ZF BioLabs will supply costumers with a digital photo of the RT-PCR gel (suitable for publication), as well as a brief description of the different stages of development in which gene expression is detected. This technique can also be used as a confirmation method for mRNA alternative splicing block of specific genes by antisense oligo (morpholinos) and its consequently loss of function (see Functional genomic studies)