---

This volume presents detailed laboratory procedures in an easy to follow format that can be carried out with success by investigators lacking previous exposure to a specific research method. Chapter guide readers through the application of molecular approaches to disease gene identification and overviews, and case studies are also presented. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Disease Gene Identification: Methods and Protocols, Second Edition aims to help with the identification and characterization of many more disease-related genes and provide novel, and effective strategies for disease treatment and prevention.

---

Part I: Introduction

1. Identification of Disease Susceptibility Alleles in the Next Generation Sequencing Era

Johanna K. DiStefano and Christopher B. Kingsley

2. Induced Pluripotent Stem Cells in Disease Modeling and Gene Identification

Satish Kumar, John Blangero, and Joanne E. Curran

3. Development of Targeted Therapies Based on Gene Modification

Taylor Benson, Fatjon Leti, and Johanna K. DiStefano

4. What can we Learn about Human Disease from the Nematode C. elegans?

Javier Apfeld and Scott AlperstParagr

5. Microbiome Sequencing Methods for Studying Human Diseases

Rebecca M. Davidson and L. Elaine Epperson

6. The Emerging Role of Long Noncoding RNAs in Human Disease

Johanna K. DiStefano

Part II: Methods for Gene Identification

7. Identification of Disease-related Genes using a Genome-wide Association Study Approach

Tobias Wohland and Dorit Schleinitz

8. Whole Genome Library Construction for Next Generation Sequencing

Jonathan
J. Keats, Lori Cuyugan, Jonathan Adkins, and Winnie S. Liang

9. Whole Exome Library Construction for Next Generation Sequencing

Winnie S. Liang, Kristi Stephenson, Jonathan Adkins, Austin Christofferson, Adrienne Helland, Lori Cuyugan, and Jonathan J. Keats

10. Optimized Methodology for the Generation of RNA-sequencing Libraries from Low-input Starting Material: Enabling Analysis of Specialized Cell Types and Clinical Samples

Brian O´Connor and Kendra Walton

11. Using C1 to Generate

Single-cell Full-length cDNA Libraries for mRNA Sequencing

Robert Durruthy-Durruthy and Manisha Ray

12. MiSeq: A Next Generation Sequencing Platform for Genomic Analysis

Rupesh Kanchi Ravi, Kendra Walton, and M
ahdieh Khosroheidari

13 Methods for CpG Methylation Array Profiling via Bisulfite Conversion

MsoListParagraphCxSpLast" style="margin-left: 0.0in;"Fatjon Leti, Lorida Llaci, Ivana Malenica, and Johanna K. DiStefano

Part III: Functional Characterization of Susceptibility Alleles and Loci

14. miRNA Quantification Me
thod using Quantitative Polymerase Chain Reaction in Conjunction with the Cq Methodspan

Fatjon Leti and Johanna K. DiStefanoserif in-left: 0.0in;"Gene Editing of Primary Human Airway Epithelial Cells

Jamie L. Everman, Cydney Rios, and Max A. Seibold

16. RNA Interference to Knockdown Gene Expression
Haiyong Han

17. Using Luciferase Reporter Assays to Identify Functional Variants at Disease-associated Loci

Anup K. Nair and Leslie J. Baier

PART IV: Identification of Disease Genes

18. Physiologic Interpretation of GWAS Signals for type 2 Diabetes

Richard M. Watanabe

19. Identification of Genes for Hereditary Hemochromatosis

Glenn S. Gerhard, Barbara V. Paynton, and Johanna K. DiStefano

20. Identification of Driver Mutations in Rare Cancers: The Role of SMARCA4 in Small Cell Carcinoma of the Ovary, Hypercalcemic Type (SCCOHT)

Jessica D. Lang and William P.D. Hendricksan

21. The Rise and Fall and Rise of Linkage Analysis as a Technique for Finding and Characterizing Inherited Influences on Disease Expression

Ettie M. Lipner and David A. Greenberg

---