QuantiLyse: Reliable DNA Amplification from Single …

QuantiLyse : Reliable DNA Amplification from Single Cells Kenneth E. Pierce, John E. Rice, J. Aquiles Sanchez, and Lawrence J. Wangh Brandeis University, Waltham, MA, USA. BioTechniques 32:1106-1111 (May 2002). ABSTRACT of PCR-based assays in both scientific investigations and clin- ical diagnosis. Although the introduction of real-time PCR in Amplification of DNA sequences from Single cells via PCR is in- the last few years has improved the quantitative rigor of such creasingly used in basic research and clinical diagnostics but re- assays, their reliability at the Single -cell level remains prob- mains technically difficult. We have developed a cell lysis protocol lematic. Such sensitivity requires that both sample prepara- that uses an optimized proteinase K solution, named QuantiLyse , tion and PCR components be thoroughly optimized. Primer and permits Reliable Amplification from individual cells.

account for the wide range of ADO rates reported in different studies (10,14,17). However, it is difficult to determine the best lysis methods from these reports, since virtually all …

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