title: A useful protocol for analyses of mutations of the epidermal growth factor receptor gene.
abstract: Recent studies have reported that mutations of the epidermal growth factor receptor (EGFR) gene are associated with the responsiveness of tyrosine kinase inhibitors (TKIs), which are molecular targets for non-small cell lung cancer (NSCLC). To provide genetic analyses for NSCLC patients, a simple and reliable method using paraffin-embedded materials is needed. The DEXPAT DNA extraction kit was used for DNA extraction from paraffin-embedded materials. DNA was amplified using the nested PCR technique, then analyzed by direct sequencing for EGFR mutations (exons 18 to 21). The phenol/chloroform extraction for DNA was also performed for comparison. When the DEXPAT kit was used, distinct bands were observed in all products after nested PCR assays of paraffin-embedded materials. Distinct sequencing signals were obtained. Results from the sequencing analysis of paraffin-embedded materials and frozen materials were completely concordant. The current study suggests that DNA extraction with the DEXPAT kit followed by nested PCR is a simple and reliable technique for analyzing the EGFR gene status with paraffin-embedded samples.
authors: Second Department of Surgery, Kagawa University, Kita-gun, Kagawa 761-0793, Japan.
authors: Dage
authors: D
authors: Liu
authors: Jun
authors: J
authors: Nakano
authors: Masaki
authors: M
authors: Ueno
authors: Daiki
authors: D
authors: Masuya
authors: Takashi
authors: T
authors: Nakashima
authors: Hiroyasu
authors: H
authors: Yokomise
authors: Kouichi
authors: K
authors: Yube
authors: Cheng-Long
authors: CL
authors: Huang