Assignment Task: Metaphase chromosomes were obtained from stimulated peripheral blood cultures. Harvesting, chromosome preparations and G-banded chromosome analysis were performed using standard techniques. For fluorescence in situ hybridization, (FISH), slides were denatured in 70% formamide at 73 C followed by sequential dehydration in 70, 85 and 100% ethanol. The slides were counterstained with 4,6-diamidino-2- phenylindole dihydrochloride (DAPI) and evaluated using a Leica DMLB fluorescent microscope.
Q1. What is the advantage of karyotyping with G-staining of chromosomes as opposed to simple karyotyping?
Q2. What does stimulated cell culture mean and why is cell stimulation performed?
Q3. What are the features of the FISH method in contrast to karyotyping with G- staining of chromosomes?
Q4. What is DNA Denaturation? Is DNA denaturation a mandatory procedure for the FISH method? Why?