A Novel DHPLC-Based Procedure for the Analysis of COL1A1 and COL1A2 Mutations in Osteogenesis Imperfecta Antonella Fuccio, Mariangela Iorio, Felice Amato,

Slides:

Advertisements

Similar presentations

Measurement of Relative Copy Number of CDKN2A/ARF and CDKN2B in Bladder Cancer by Real-Time Quantitative PCR and Multiplex Ligation-Dependent Probe Amplification.

Advertisements

CyclinD1/CyclinD3 Ratio by Real-Time PCR Improves Specificity for the Diagnosis of Mantle Cell Lymphoma Carol D. Jones, Katherine H. Darnell, Roger A.

Design and Multiseries Validation of a Web-Based Gene Expression Assay for Predicting Breast Cancer Recurrence and Patient Survival Ryan K. Van Laar The.

Keyur P. Patel, Bedia A. Barkoh, Zhao Chen, Deqin Ma, Neelima Reddy, L

Diagnostic Testing for Rett Syndrome by DHPLC and Direct Sequencing Analysis of the MECP2 Gene: Identification of Several Novel Mutations and Polymorphisms

Simultaneous Genotyping of α-Thalassemia Deletional and Nondeletional Mutations by Real-Time PCR–Based Multicolor Melting Curve Analysis Qiuying Huang,

Mutation Spectrum of the Survival of Motor Neuron 1 and Functional Analysis of Variants in Chinese Spinal Muscular Atrophy Yu-jin Qu, Jin-li Bai, Yan-yan.

A Multiplex SNaPshot Assay as a Rapid Method for Detecting KRAS and BRAF Mutations in Advanced Colorectal Cancers Sandrine Magnin, Erika Viel, Alice.

Β-Glucuronidase Is an Optimal Normalization Control Gene for Molecular Monitoring of Chronic Myelogenous Leukemia Joong Won Lee, Qiaofang Chen, Daniel.

Wanlong Ma, Hagop Kantarjian, Xi Zhang, Chen-Hsiung Yeh, Zhong J

Mutation Screening in Juvenile Polyposis Syndrome

Betaine, Dimethyl Sulfoxide, and 7-Deaza-dGTP, a Powerful Mixture for Amplification of GC-Rich DNA Sequences Marco Musso, Renata Bocciardi, Sara Parodi,

Joanna Wang, Chetan Bettegowda The Journal of Molecular Diagnostics

Analysis of the COL1A1 and COL1A2 Genes by PCR Amplification and Scanning by Conformation-Sensitive Gel Electrophoresis Identifies Only COL1A1 Mutations.

Philippe Szankasi, Mohamed Jama, David W. Bahler

Comparison of Allelic Discrimination by dHPLC, HRM, and TaqMan in the Detection of BRAF Mutation V600E Pablo Carbonell, María C. Turpin, Daniel Torres-Moreno,

Quantitative Measurement of Cell-Free Plasma DNA and Applications for Detecting Tumor Genetic Variation and Promoter Methylation in a Clinical Setting

Molecular Diagnosis in Ewing Family Tumors

Undifferentiated Small Round Cell Sarcomas with Rare EWS Gene Fusions

Simultaneous Genotyping of α-Thalassemia Deletional and Nondeletional Mutations by Real-Time PCR–Based Multicolor Melting Curve Analysis Qiuying Huang,

Thomas W. Prior, Scott J. Bridgeman

Getting Things Backwards to Prevent Primer Dimers

Patrick R. Murray The Journal of Molecular Diagnostics

Detection of an Apparent Homozygous 3120G>A Cystic Fibrosis Mutation on a Routine Carrier Screen Denise LaMarche Heaney, Patrick Flume, Lauren Hamilton,

Andrea Gaedigk, Amanda K. Riffel, J. Steven Leeder

Low Copy Number DNA Template Can Render Polymerase Chain Reaction Error Prone in a Sequence-Dependent Manner Mansour Akbari, Marianne Doré Hansen, Jostein.

Hillary S. Sloane, James P. Landers, Kimberly A. Kelly

Pathogenicity Evaluation of BRCA1 and BRCA2 Unclassified Variants Identified in Portuguese Breast/Ovarian Cancer Families Catarina Santos, Ana Peixoto,

Catherine E. Keegan, Anthony A. Killeen

Rapid Detection of TEM-Type Extended-Spectrum β-Lactamase (ESBL) Mutations Using Lights-On/Lights-Off Probes with Single-Stranded DNA Amplification Kenneth.

Sensitive Detection of Deletions of One or More Exons in the Neurofibromatosis Type 2 (NF2) Gene by Multiplexed Gene Dosage Polymerase Chain Reaction

Double Heterozygosity for a RET Substitution Interfering with Splicing and an EDNRB Missense Mutation in Hirschsprung Disease Alberto Auricchio, Paola.

Harry R. Hill, Nancy H. Augustine, Robert J. Pryor, Gudrun H

Molecular Characterization of WFS1 in Patients with Wolfram Syndrome

Alexandra Scott, Hanna M

Analysis of Rare APC Variants at the mRNA Level

Keyur P. Patel, Bedia A. Barkoh, Zhao Chen, Deqin Ma, Neelima Reddy, L

The c.1364C>A (p.A455E) Mutation in the CFTR Pseudogene Results in an Incorrectly Assigned Carrier Status by a Commonly Used Screening Platform Kristin.

Molecular and Functional Analysis of the Large 5′ Promoter Region of CFTR Gene Revealed Pathogenic Mutations in CF and CFTR-Related Disorders Sonia Giordano,

Benjamin P. Song, Surbhi Jain, Selena Y. Lin, Quan Chen, Timothy M

BRAF Mutation Testing in Solid Tumors

Skin Biopsy Is a Practical Approach for the Clinical Diagnosis and Molecular Genetic Analysis of X-Linked Alport's Syndrome Fang Wang, Dan Zhao, Jie.

A Synonymous Mutation in the CFTR Gene Causes Aberrant Splicing in an Italian Patient Affected by a Mild Form of Cystic Fibrosis Valeria Faa′, Alessandra.

Iris Schrijver, Tiffanee J. Lenzi, Carol D. Jones, Marla J

WGA Allows the Molecular Characterization of a Novel Large CFTR Rearrangement in a Black South African Cystic Fibrosis Patient Marie des Georges, Caroline.

A Highly Sensitive Genetic Protocol to Detect NF1 Mutations

Evaluating the Effect of Unclassified Variants Identified in MMR Genes Using Phenotypic Features, Bioinformatics Prediction, and RNA Assays Lucia Pérez-Cabornero,

Sarah E. Kerr, Cheryl B. Thomas, Stephen N. Thibodeau, Matthew J

A Novel Long-Range PCR Sequencing Method for Genetic Analysis of the Entire PKD1 Gene Ying-Cai Tan, Alber Michaeel, Jon Blumenfeld, Stephanie Donahue,

Ye Bang-Ce, Chu Xiaohe, Fan Ye, Li Songyang, Yin Bincheng, Zuo Peng

A Novel Method for Creating Artificial Mutant Samples for Performance Evaluation and Quality Control in Clinical Molecular Genetics Michael Jarvis, Ramaswamy.

Development of a High-Resolution Melting Curve Analysis Screening Test for SRSF2 Splicing Factor Gene Mutations in Myelodysplastic Syndromes Eduardo.

Validation of Denaturing High Performance Liquid Chromatography as a Rapid Detection Method for the Identification of Human INK4A Gene Mutations Irene.

Validation of High-Resolution DNA Melting Analysis for Mutation Scanning of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Gene Marie-Pierre.

Feras M. Hantash, Arlene Rebuyon, Mei Peng, Joy B

Danielle C. Smith, Alina Esterhuizen, Jacquie Greenberg

Rapid Polymerase Chain Reaction-Based Detection of Epidermal Growth Factor Receptor Gene Mutations in Lung Adenocarcinomas Qiulu Pan, William Pao, Marc.

Improved Detection of KIT Exon 11 Duplications in Formalin-Fixed, Paraffin-Embedded Gastrointestinal Stromal Tumors Jerzy Lasota, Bartosz Wasag, Sonja.

A Novel and Rapid Method of Determining the Effect of Unclassified MLH1 Genetic Variants on Differential Allelic Expression Sheron Perera, Brian Li,

Assessing the Functional Characteristics of Synonymous and Nonsynonymous Mutation Candidates by Use of Large DNA Constructs A.M. Eeds, D. Mortlock, R.

Two Novel Methods for Rapid Detection and Quantification of DNMT3A R882 Mutations in Acute Myeloid Leukemia Melissa Mancini, Syed Khizer Hasan, Tiziana.

Wook Lew Journal of Investigative Dermatology

Extensive Molecular Analysis of Patients Bearing CFTR-Related Disorders Felice Amato, Chiara Bellia, Giuseppe Cardillo, Giuseppe Castaldo, Marcello Ciaccio,

KIT Gene Deletions at the Intron 10−Exon 11 Boundary in GI Stromal Tumors Christopher L. Corless, Laura McGreevey, Ajia Town, Arin Schroeder, Troy Bainbridge,

Extra Alleles in FMR1 Triple-Primed PCR

Xiangfeng Cui, Helen Feiner, Honghua Li

Improved Detection of the KIT D816V Mutation in Patients with Systemic Mastocytosis Using a Quantitative and Highly Sensitive Real-Time qPCR Assay Thomas.

A Second Leaky Splice-Site Mutation in the Spastin Gene

Optimized Allele-Specific Real-Time PCR Assays for the Detection of Common Mutations in KRAS and BRAF Alois H. Lang, Heinz Drexel, Simone Geller-Rhomberg,

Exon Skipping in IVD RNA Processing in Isovaleric Acidemia Caused by Point Mutations in the Coding Region of the IVD Gene Jerry Vockley, Peter K. Rogan,

Presentation transcript:

A Novel DHPLC-Based Procedure for the Analysis of COL1A1 and COL1A2 Mutations in Osteogenesis Imperfecta Antonella Fuccio, Mariangela Iorio, Felice Amato, Ausilia Elce, Rosaria Ingino, Mirella Filocamo, Giuseppe Castaldo, Francesco Salvatore, Rossella Tomaiuolo The Journal of Molecular Diagnostics Volume 13, Issue 6, Pages 648-656 (November 2011) DOI: 10.1016/j.jmoldx.2011.06.006 Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 DHPLC profiles obtained in a patient with OI bearing a mutation in COL1A1 (profile A) and in a control individual bearing a variant in COL1A1 when testing DNA from two cells of buccal mucosa (profile B). The Journal of Molecular Diagnostics 2011 13, 648-656DOI: (10.1016/j.jmoldx.2011.06.006) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 A: Schematic structure of the pEGFP-C3-Col1A(1-2) minigene constructs and position of the unclassified variants. Green fluorescent protein and COL1A1 or COL1A2 exons are shown as large boxes, and the lines between exons represent the corresponding introns. The thin box indicates the CMV promoter (pCMV), and the black arrows represent the location of the primers used for the splicing pattern analysis. Also shown are alternative predicted splicing donor (d) and acceptor (a) sites, as well as classic splicing donor (D) and acceptor (A) sites. B–D: Splicing pattern analysis of the pEGFP-C3-Col1A(1-2) minigene constructs carrying different genetic variants (M) are compared with their relative wild-type (W) form. The structures of the PCR products are schematically shown to the right of each gel. B: The c.804 + 2_804 + 3delTG variant causes complete failure of the normal splicing pattern, which is replaced by a new splicing pattern with the presence of a predominant form that retains intron 11 and a less representative form in which the presence of an alternative donor (d) site into exon 10 causes the alternative splicing of exon 11 and part of exon 10. C and D: The variants c.3046-4_3046-5dupCT in COL1A1 and c.891 + 77A>T in COL1A2 do not affect splicing. The Journal of Molecular Diagnostics 2011 13, 648-656DOI: (10.1016/j.jmoldx.2011.06.006) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions