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Direct polymerase chain reaction amplification of formalin-fixed, paraffin-wax-embedded tissue after automated sequential laser microdissection and pressure catapulting
  1. S L O’Kane,
  2. V Garimella*,
  3. N Sivarajasingham*,
  4. P J Drew,
  5. L Cawkwell
  1. Cancer Biology Proteomics Group, Postgraduate Medical Institute of the University of Hull in association with the Hull-York Medical School, University of Hull, Hull, UK
  1. Correspondence to:
 Dr L Cawkwell
 R&D Building, Castle Hill Hospital, Hull, HU16 5JQ, UK; l.cawkwell{at}hull.ac.uk

Abstract

A robust method to facilitate rapid laser microdissection and pressure catapulting (LMPC) coupled with direct polymerase chain reaction (dPCR) to eliminate the need for extraction of DNA before a PCR-based assay is described. This sequential LMPC–dPCR method is rapid and decreases the number of processing steps, reducing the chance of tissue loss and contamination.

  • dPCR, direct polymerase chain reaction
  • FFPE, formalin-fixed, paraffin-wax-embedded
  • LMPC, laser microdissection and pressure catapulting
  • PCR, polymerase chain reaction
  • PEN, polyethylene naphtahalate
  • sLMPC, sequential laser microdissection and pressure catapulting

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Footnotes

  • * These authors have contributed equally.

  • Competing interests: None.

  • Ethical approval: This work is covered by Hull & East Yorkshire Local Research Ethics Committee approval 04/Q1104/24.

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