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This volume provides a thorough guide of the study of multiple myeloma (MM)-from sample acquisition to the performance of molecular tests, and also discusses difficulties that arise during quality control and interpretation of results. The chapters in this book cover topics, such as next-generation-flow-based methods for detection of minimal residual disease and circulating tumor cells; Cytometry by Time-of-Flights (CyTOF); a droplet digital PCR approach for finding contamination in patients´ samples; genome-wide investigation of the methylation and hydroxymethylation status of cytosines; and a microfluidic approach for creating nanoparticles. 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.

Cutting-edge and comprehensive, Multiple Myeloma: Methods and Protocols is a valuable resource for molecular scientists and laboratory supervisors working with MM samples, and any investigator conducting studies in this field.

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Preface...

Table of Contents...

Contributing Authors...

1. ELDA qASO-PCR for High Sensitivity Detection of Tumor Cells in Bone Marrow and Peripheral Blood

Stefanie Huhn

2. EuroFlow-Based Next-Generation Flow Cytometry for Detection of Circulating Tumor Cells and Minimal Residual Disease in Multiple Myeloma

Leire Burgos Rodriguez and Bruno Paiva

3. Cytoplasmic Immunoglobulin vs DNA Analysis by Flow Cytometry

Daisy Alapat

4. Deep Profiling of the Immune System of Multiple Myeloma Patients by Cytometry by Time of Flight (CyTOF)

Tina Smets, Frederik Stevenaert, Homer Adams III, and Greet Vanhoof

5. Fluorescence In Situ Hybridization (FISH) in Multiple Myeloma

Erming Tian

6. Whole Exome Sequencing in Multiple Myeloma to Identify Somatic Single Nucleotide Variants and Key Translocations Involving Immunoglobulin Loci and MYC

Brian A. Walker

7. RNA-Sequencing from Low-Input Material in Multiple Myeloma for Application in Clinical Routine

Anja Seckinger, Tomi Bähr-Ivacevic, Vladimir Benes, and Dirk Hose

8. Protocol for M3P: A Comprehensive and Clinical Oriented Targeted Sequencing Panel for Routine Molecular Analysis in Multiple Myeloma

Santiago Barrio, Matteo Da Via, Laura Bruins, Thorsten Stühmer, Torsten Steinbrunn, Max Bittrich, Hermann Einsele, Alexander Keith stewart, Esteban Braggio, and Klaus Martin Kortüm

9. Analysis of Circulating Tumor DNA

Sridurga Mithraprabhu, and Andrew Spencer

10. Detection of Cross-Sample Contamination in Multiple Myeloma Samples and Sequencing Data

Owen W. Stephens, Tobias Meißner, and Niels Weinhold

11. Analysis of Global Gene Expression Profiles

Alboukadel Kassambara and Jerome Moreaux

12. Genome Wide Mapping of Methylated and Hydroxyl-Methylated Cytosines using a Modified HpaII Tiny Fragment Enrichment by Ligation Mediated PCR Tagged Sequencing Protocol

Sanchari Battachariyya, Ruslana Tytarenko, Christoph Heuck, John Greally, and Amit Verma

13. A Rapid and Robust Protocol for Reduced Representation Bisulfite Sequencing in Multiple Myeloma

Samrat Roy Choudhury and Brian A. Walker

14. Microfluidic Production and Application of Lipid Nanoparticles for Nucleic Acid Transfection

Anitha Thomas, Shyam Garg, Rebecca DeSousa, Eric Ouellet, Grace Tharmarajah, Dave Reichert, Mina Ordobadi, Shell Ip, Samuel Clarke, and Euan C. Ramsay

15. Microfluidic Assembly of Liposomes with Tunable Size and Co-Loading Capabilities

Jessica R. Hoffman, Ennio Tasciotti, and Roberto Molinaro

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