Novel strategies to clone identical and distinct DNA sequences for several complex genomes
Tóm tắt
In this minireview I briefly describe the new methods suggested for cloning sequences identical by descent, homo-or hemizygously deleted, amplified or polymorphic, and compare them with the most efficient techniques developed earlier. The new methods include cloning of identical sequences (CIS), cloning of polymorphic sequences (COP), and cloning of deleted sequences (CODE). Although these methods are based on the same combination of biochemical techniques, their aims are different. These methods are fully complementary, and they may be combined to analyze a given object. If one aims to clone a disease gene responsible for familial cancer syndrome, these methods may be applied as follows. CIS can be used to identify the sequences identical by descent comparing the DNA obtained from affected or unaffected family members. COP can be used to find sequences that are different between affected and unaffected members, and CODE would be useful to compare tumor and normal (control) samples to isolate, deleted sequences (putative candidate tumor suppressor genes) and amplified sequences (putative oncogenes). The COP and CODE procedures can be applied to analyze the CpG islands, thus allowing direct candidate gene identification.
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