Fluorescence In Situ Hybridization Pnafish

The efficiency of PNAs as hybridization probes has also been demonstrated in fluorescence in situ hybridization

Fig. 2 In situ identification of chromosomes 1, X, and Y on human metaphase and interphase nuclei using multicolor PNA-FISH procedure with satellite-specific PNA probes. Chromosome 1 is labeled in blue, chromosome X in red, and chromosome Y in green. (View this art in color at www. dekker.com.)

polymorphisms. Recently, PNA-FISH has been adapted to in situ chromosomal analysis of the human sperm.[22] This adaptation constituted an interesting challenge because of the particularities of human spermatozoa nuclei in terms of genomic compaction and accessibility of DNA sequences. Comparative estimates of disomies X, Y, and 1 were performed in sperm from healthy subjects using FISH, primed in situ labeling (PRINS), and PNA procedures in parallel. Equivalent quality of in situ nuclear labeling and similar results was obtained with the three methods. However, the hybridization timing of PNA probes (i.e., 45 min) appeared to be significantly shortened in comparison with FISH reaction on sperm. The fast hybridization kinetics of PNA was similar to the kinetics of PRINS reaction.[23] The similarity between PNA and PRINS might be essentially due to the small size of both PNA probes and PRINS primers, which do not exceed 30 bases in length. This finding points out the importance of the probe size for in situ assays and, consequently, the great potentiality of the PNA approach, because high specific binding can be obtained with a short, unique PNA oligomer. This study also demonstrates the efficiency of multicolor PNA procedure on male gametes and shows that PNA could be a powerful alternative to FISH and PRINS for in situ chromosomal investigations.

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

Get My Free Ebook


Post a comment