Development of a Prognostic Marker for Lung Cancer Using Analysis of Tumor Evolution
Technical Report,01 Aug 2015,31 Jul 2016
Duke University Durham United States
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The goal of this project is to sequence the exomes of single tumor cells from tumors in order to construct evolutionary trees, the characteristics of which will be used to predict whether a tumor will metastasize or not. We established a procedure for isolating tumor cells from lung tumor tissue and observed high variability in yield. We also encountered loss of tumor cell viability after freezing. Despite these challenges, we will be able to obtain sufficient numbers of cells to proceed. The next step is whole genome amplification of the DNA from single cells as a prerequisite to sequencing. We encountered a low success rate of whole genome amplification from single sorted cells and have spent a great deal of time troubleshooting this process. We anticipate that exome sequencing of single tumor cells should begin shortly in order to reconstruct tumor cell evolution in tumors.
- Medicine and Medical Research
- Genetic Engineering and Molecular Biology