We create a novel approach that is non-destructive towards the embryo for molecular sex recognition of embryonic specimens. Embryonic bloodstream from inside regarding the eggshell ended up being swabbed onto a FTA ® Elute Micro Card (Whatman) just after egg dissection. DNA had been extracted after the manufacturer’s directions with a protocol adapted for automatic analysis that is high-throughput the Eppendorf EPmotion 5075 liqu >® card extractions of adult P. vitticeps bloodstream examples (letter = 30).
We then carried out a PCR-based test, which will be diagnostic when it comes to existence for the W chromosome. PCR conditions observed Holleley et al. 14; however, as a result of the probability of low DNA levels from embryonic product, we increased the quantity of DNA put into PCRs (3 µl per effect; around 65 ng DNA per PCR). Continue reading