PCR-free quantification of multiple splice variants in a cancer gene by surface-enhanced raman spectroscopy

Research output: Contribution to journalArticle

Abstract

We demonstrate a surface-enhanced Raman spectroscopy (SERS) based array platform to monitor gene expression in cancer cells in a multiplex and quantitative format without amplification steps. A strategy comprising DNA/RNA hybridization, S1 nuclease digestion, and alkaline hydrolysis was adopted to obtain DNA targets specific to two splice junction variants, δ(9,10) and A(5), of the breast cancer susceptibility gene 1 from MCF-7 and MDA-MB-231 breast cancer cell lines. These two targets were identified simultaneously, and their absolute quantities were estimated by a SERS strategy utilizing the inherent plasmon-phonon Raman mode of gold nanoparticle probes as a self-referencing standard to correct for the variability in surface enhancement. The results were then validated by reverse-transcription polymerase chain reaction. Our proposed methodology could be expanded to a higher level of multiplexing for quantitative gene expression analysis of any gene without any amplification steps.

Original languageEnglish (US)
Pages (from-to)14021-14025
Number of pages5
JournalJournal of Physical Chemistry B
Volume113
Issue number42
DOIs
StatePublished - Oct 22 2009
Externally publishedYes

Fingerprint

oncogenes
Raman Spectrum Analysis
Neoplasm Genes
Raman spectroscopy
Genes
gene expression
cancer
Phonons
Breast Neoplasms
Gene Expression
Gene expression
breast
genes
Polymerase Chain Reaction
Amplification
DNA
deoxyribonucleic acid
Cells
Gold
Nanoparticles

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Surfaces, Coatings and Films
  • Materials Chemistry

Cite this

PCR-free quantification of multiple splice variants in a cancer gene by surface-enhanced raman spectroscopy. / Sun, Lan; Irudayaraj, Joseph.

In: Journal of Physical Chemistry B, Vol. 113, No. 42, 22.10.2009, p. 14021-14025.

Research output: Contribution to journalArticle

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