Application of optical trapping for cells grown on plates: Optimization of PCR and fidelity of DNA sequencing of p53 gene from a single cell

From: Clinical Chemistry | Date: March 1, 2003| Author: | Copyright information

Background: Optical trapping has traditionally been used to visually select and isolate nonadherent cells grown in suspension because cells grown in monolayers will rapidly reattach to surfaces if suspended in solution. We explored methods to slow cell reattachment that are also compatible with high-fidelity PCR.

Methods: Using HeLa cells grown on plates and suspended after trypsinization, we measured the efficiency of capture by retention and movement of the cell by the laser. Success for removing a captured cell by pipette was determined by PCR amplification of the 5S rRNA gene. After ...

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