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Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Vienna experience

Leukemia volume 17, pages 224–227 (2003)Cite this article

The method is based on STR (short tandem repeat)-PCR analysis1,2,3,4,5,6,7,8,9,10,11using fluorescence-labelled primers. The characteristics of the polymorphic markers tested are presented in Table 1. PCR products are analyzed and quantified by a capillary electrophoresis instrument (ABI Prism 310 Genetic Analyzer). Calculation of donor chimerism is based on the ratio of informative donor and recipient peaks.10

Table 1 Characteristics of the marker panel
Full size table

Initial genotyping for the detection of informative STR loci is usually performed using peripheral blood (PB) from the recipient and PB or bone marrow (BM) from the donor. The Qiagen DNA Blood Kit (Qiagen, Hilden, Germany) is used according to the manufacturer's recommendations. The DNA yield is quantified by spectrophotometry and 10 ng of DNA are used as template in individual PCR reactions.

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Acknowledgements

This work was supported by the ‘Österreichische Kinderkrebshilfe’.

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  1. Children's Cancer Research Institute, Vienna, Austria

    E Schraml, H Daxberger, F Watzinger & T Lion

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Schraml, E., Daxberger, H., Watzinger, F. et al. Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Vienna experience. Leukemia 17, 224–227 (2003). https://doi.org/10.1038/sj.leu.2402756

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