Product Description:
The general principal behind in situ hybridization involves specific annealing of labeled DNA probe to complementary sequence(s) in fixed specimen that is followed by visualization of the location of the probe. Since the late 1980s fluorescence in situ hybridization (FISH) has evolved into a useful tool for precise and relatively prompt results. More recently, FISH is used as a confirmatory technique for various deletions/duplications detected by array comparative genomic hybridization (aCGH). Home-brew FISH probes are developed for these verification studies. The entire method from a target sequence selection to growth, extractions, labeling, hybridization, and detection presents time challenges, especially when being performed as confirmatory studies following aCGH investigation. To achieve rapid results using home-brew probes, our laboratory has modified a time saving protocol on cultured blood specimens. The feasibility of this protocol with other types of specimens is yet to be tested. Normally, a conventional home-brew FISH probe development from bacterial artificial clones (BAC) entails seven to nine days; however our modified procedure plates the clone (day one), liquid inoculation (day two), then extraction and labeling (day three), while the precipitation, rapid hybridization, and probe analysis is combined into one day (day four) allotting a swift turn around time for probe development. Fifteen FISH proves were developed using the modified protocol. The procedure was successfully completed in four to five days in all 15 (100%) studies. Our results demonstrated that our methods can be quite effective with a time-frame of four to five days. The rapid hybridization procedure was successful for 12/15 probes (80%), while 3/15 (20%) endured an overnight hybridization from a failed rapid hybridization to affirm appropriate results. We provide a modified protocol which is beneficial when facing time restraints. The technique does not entail any extra equipment or chemicals. Our protocol is thus cost-effective as well as time-saving and serves to facilitate rapid and accurate results without compromising the quality.
