Optimization of a multiplex minisequencing protocol for population studies and medical genetics

Cláudia M B Carvalho, Sérgio D J Pena
<span title="2005-06-30">2005</span> <i title="Genetics and Molecular Research"> <a target="_blank" rel="noopener" href="https://fatcat.wiki/container/tf4txbjapvcg5ds7uwx3x2vhnm" style="color: black;">Genetics and Molecular Research</a> </i> &nbsp;
Several technologically sophisticated high-throughput techniques have been recently developed for the study of human single nucleotide polymorphisms and the diagnosis of point mutations in human diseases. However, there is also a need for simple and inexpensive techniques suitable for clinical services and small research laboratories. Minisequencing meets the latter requirements. It is simple, non-radioactive and can be easily multiplexed by adding oligonucleotide tails of increasing size to
more &raquo; ... sequencing oligonucleotide primers. To optimize the minisequencing protocol, we designed a test multiplex system capable of typing simultaneously 12 different human autosomal single nucleotide polymorphisms. We discovered that the quality of minisequencing primers and the careful selection of the tail sequences were especially critical for success. This optimized protocol permits rapid genotyping at low cost and can serve as a blueprint for the creation of multiplex minisequencing systems suitable to virtually any typing application in population studies and medical genetics.
<span class="external-identifiers"> <a target="_blank" rel="external noopener" href="https://www.ncbi.nlm.nih.gov/pubmed/16110434">pmid:16110434</a> <a target="_blank" rel="external noopener" href="https://fatcat.wiki/release/na6xl4v5vjbvnmzs2ynb7ifvnu">fatcat:na6xl4v5vjbvnmzs2ynb7ifvnu</a> </span>
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