BACKGROUND: The use of paraffin wax embedded tissue samples as a source of DNA for genotype analysis has been limited because of difficulties in DNA extraction and single nucleotide polymorphism (SNP) analysis. AIMS: To test the feasibility of applying the combination of a commonly used DNA isolation procedure, PureGene, and a high throughput SNP analysis method, the polymerase chain reaction (PCR)-INVADER assay, to genotype several types of paraffin wax embedded breast tissues. METHODS: Twenty formalin fixed, paraffin wax blocks were obtained from five participants in the Iowa women's health study. Each participant provided several types of tissue including normal lymph node, normal nipple/areola tissue, inflammatory/fibrotic breast tissue, or normal breast tissue, and tumour tissue. RESULTS: Good quality DNA (260/280 ratio >1.6) was obtained from all tissues. Normal lymph nodes yielded the largest amount of DNA (97.1 mug). DNA obtained from the samples was tested for a germline C1183T polymorphism in the MnSOD gene by three methods-PCR-RFLP (restriction fragment length polymorphism), INVADER assay, and PCR-INVADER assay. Of the 20 samples, PCR-RFLP genotyped 16, the PCR-INVADER assay 18, and the INVADER assay two. This methodology was then used to analyse five additional genotypes and confirmed the general applicability of the method. CONCLUSIONS: This study demonstrated the feasibility of (1) using several paraffin wax embedded breast tissues as a source of DNA for germline genetic analysis, with lymph nodes providing the highest yield, and (2) using the combination of a common extraction method with a high throughput SNP analysis method, the PCR-INVADER assay.
BACKGROUND: The use of paraffin wax embedded tissue samples as a source of DNA for genotype analysis has been limited because of difficulties in DNA extraction and single nucleotide polymorphism (SNP) analysis. AIMS: To test the feasibility of applying the combination of a commonly used DNA isolation procedure, PureGene, and a high throughput SNP analysis method, the polymerase chain reaction (PCR)-INVADER assay, to genotype several types of paraffin wax embedded breast tissues. METHODS: Twenty formalin fixed, paraffin wax blocks were obtained from five participants in the Iowa women's health study. Each participant provided several types of tissue including normal lymph node, normal nipple/areola tissue, inflammatory/fibrotic breast tissue, or normal breast tissue, and tumour tissue. RESULTS: Good quality DNA (260/280 ratio >1.6) was obtained from all tissues. Normal lymph nodes yielded the largest amount of DNA (97.1 mug). DNA obtained from the samples was tested for a germline C1183T polymorphism in the MnSOD gene by three methods-PCR-RFLP (restriction fragment length polymorphism), INVADER assay, and PCR-INVADER assay. Of the 20 samples, PCR-RFLP genotyped 16, the PCR-INVADER assay 18, and the INVADER assay two. This methodology was then used to analyse five additional genotypes and confirmed the general applicability of the method. CONCLUSIONS: This study demonstrated the feasibility of (1) using several paraffin wax embedded breast tissues as a source of DNA for germline genetic analysis, with lymph nodes providing the highest yield, and (2) using the combination of a common extraction method with a high throughput SNP analysis method, the PCR-INVADER assay.
Authors: Michael Olivier; Lee-Ming Chuang; Mau-Song Chang; Ying-Tsung Chen; Dee Pei; Koustubh Ranade; Anniek de Witte; Jennifer Allen; Nguyet Tran; David Curb; Richard Pratt; Henk Neefs; Monika de Arruda Indig; Scott Law; Bruce Neri; Lu Wang; David R Cox Journal: Nucleic Acids Res Date: 2002-06-15 Impact factor: 16.971
Authors: Monika de Arruda; Victor I Lyamichev; Peggy S Eis; Walter Iszczyszyn; Robert W Kwiatkowski; Scott M Law; Marilyn C Olson; Eric B Rasmussen Journal: Expert Rev Mol Diagn Date: 2002-09 Impact factor: 5.225
Authors: R Sachidanandam; D Weissman; S C Schmidt; J M Kakol; L D Stein; G Marth; S Sherry; J C Mullikin; B J Mortimore; D L Willey; S E Hunt; C G Cole; P C Coggill; C M Rice; Z Ning; J Rogers; D R Bentley; P Y Kwok; E R Mardis; R T Yeh; B Schultz; L Cook; R Davenport; M Dante; L Fulton; L Hillier; R H Waterston; J D McPherson; B Gilman; S Schaffner; W J Van Etten; D Reich; J Higgins; M J Daly; B Blumenstiel; J Baldwin; N Stange-Thomann; M C Zody; L Linton; E S Lander; D Altshuler Journal: Nature Date: 2001-02-15 Impact factor: 49.962
Authors: P L Paris; P A Kupelian; J M Hall; T L Williams; H Levin; E A Klein; G Casey; J S Witte Journal: Cancer Epidemiol Biomarkers Prev Date: 1999-10 Impact factor: 4.254
Authors: A C Deitz; W Zheng; M A Leff; M Gross; W Q Wen; M A Doll; G H Xiao; A R Folsom; D W Hein Journal: Cancer Epidemiol Biomarkers Prev Date: 2000-09 Impact factor: 4.254
Authors: Emmanuel Amankwaa-Frempong; Francis Agyemang Yeboah; Samuel Blay Nguah; Lisa A Newman Journal: JAMA Surg Date: 2017-08-01 Impact factor: 14.766