DNeasy Blood and Tissue Kit (250)

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	Use of 5 * 106 cells to extract DNA

Protocol tips
Use of 5 * 106 cells to extract DNA

Publication protocol

Growth and isolation of cells: The cells were grown in Rose Roswell Park Memorial Institute (RPMI-1640) medium (Life Technologies, Carlsbad, USA), supplemented with 10% fetal bovine serum (Life Technologies, Carlsbad, USA) at a temperature of 37 °C and a 5% CO2 level. A known number of cells was collected with a denuding handle from STRIPPER (Origio, Måløv, Denmark) and MXL3-100 needles with a diameter of 100 µm (Origio, Måløv, Denmark) from a serial dilution of medium with phosphate buffered saline (PBS). All cells were collected in a maximum total volume of 2.5 µl and snap frozen in liquid N2 immediately after collection. For the bulk DNA sample, DNA was extracted from 5 * 106 cells using the DNeasy Blood & Tissue kit (Qiagen Hilden, Germany).

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