In the years following Donahue's work but preceding the entire sequencing of the human genome, great strides were made within the mapping and cloning of human genes; most of those genes were related to single-gene diseases. for instance, two years after Donahue used a natural action in chromosome structure to determine a link with the Duffy people locus, chromosomal banding patterns were discovered, which allowed researchers to spot deletions, insertions, and translocations that were linked to disease in pedigrees and to map the corresponding genes to particular chromosomal locations.

In addition to identifying linkage between altered chromosome structure and a given gene, researchers also recognized that there are sites in our chromosomes that may vary from one person to the subsequent. These sites are brought up as polymorphisms, and scientists were ready to develop specific DNA probes that recognize and bind to those polymorphic DNA regions. as an example, in some cases, the altered DNA sequence ends up in a change in a very restriction nuclease site, and a characteristic banding pattern occurs when a DNA probe is hybridized to the chromosomal DNA in an exceedingly Southern blot analysis; this altered pattern of bands is noted as a fragment length polymorphism (RFLP). RFLP analyses will be utilized in combination with large pedigrees to see RFLP patterns that are linked to numerous diseases, like Huntington's chorea. Those DNA probes that show a disease-associated RFLP can successively be wont to probe human-mouse cell hybrids to work out the chromosomal location of the disease-associated gene. Indeed, by 1991, the list of human disease-associated genes identified using RFLP-based approaches was impressively long.

The same DNA probes used for RFLP analyses may be utilized in a method called fluorescence in place hybridization. During this case, the probe is labeled with a fluorescent tag and incubated with a metaphase chromosome spread. The DNA probe will hybridize to its corresponding chromosomal region, and therefore the fluorescent signal will be detected and mapped.

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