Polymerase comparative sequence analysis

Michel, F., and Westhof, E. (1990). Modelling of the three-dimensional architecture of group I catalytic introns based on comparative sequence analysis. J. Mol. Biol. 216, 585-610. Milligan, J. F., and Uhlenbeck, O. C. (1989). Synthesis of small RNAs using T7 RNA polymerase. Methods Enzymol. 180, 51—62. 

In forensic cases, DNA samples can be extracted and purified from small specimens of skin, blood, semen, or hair roots collected at the crime scene. DNA that is suitable for analysis even can be obtained from dried stains of semen and blood. The RFLP analysis performed on these samples then is compared to those performed on samples obtained from the suspect. If the RFLP patterns match, it is then beyond reasonable doubt that the suspect was at the crime scene. In practice, several different probes containing different types of repetitious sequences are used in the hybridizations in order to satisfy certain statistical criteria for absolute, positive identification. The use of different restriction enzymes allow for accuracies in positive identifications of greater than one in 100 million. In recent years, polymerase chain reaction (PCR), a method that amplifies DNA, has made it possible for very small amounts of DNA found at crime scenes to be amplified for DNA fingerprinting analysis. Using specific probes to prime DNA polymerase, many copies of the targeted areas of DNA can be synthesized in vitro and subsequently analyzed. 

The analysis of DNA by CE represents one of the best examples for application of this technique in clinical analysis where the low cost, the full automation, and the speed of analysis compared to the slab gels accelerated the implementation of this technique in sequencing the human genome. Polymerase chain reaction (PCR), the Genome project, viral detection, blood transfusion safety, and forensic identification are all pushing the widespread use of DNA analysis especially by CE further than most investigators have expected. 

Expression of functional 5 -deiodinase is surprisingly low in the thyroid tissue and cells of the omnivorous pig (similar to the herbivores) compared to other omnivores such as man, rodents, and guinea pigs (Beech etal. 1993). KOhrle could not even detect steady-state mRNA levels by Northern blotting analysis or reverse transcriptase polymerase chain reaction using primer pairs complementary to the rat or human 5 -deiodinase sequence. 

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