Next Generation Sequencing (NGS) has revolutionized the way that we address complex biological questions. As sequencing output rapidly increases and experimental scales get bigger, library preparation becomes one of the major bottlenecks for NGS. This course provides a comprehensive hands-on training on how to prepare high quality libraries for Illumina HiSeq2500 and MiSeq sequencing platforms. Participants are encouraged to bring their own total RNA template and, by the end of the course, will have a library ready for NGS sequencing. Lectures will cover the entire workflow including sample QC/QA, as well as the basic principles of NGS technology and consideration for experimental design meeting current publication standards. The course will highlight applications such as mRNA-seq, ChIP-seq, de novo sequencing, re-sequencing, mutation discovery, single cell genomics, and PacBio RSII. A case example of combinatorial usage of NGS technology to meet specific biological goals of sequencing will be explored.
Protocols covered in this course are compatible with multiplexing as well as with the preparation of PCR-free, ribosomal RNA depleted, and strand specific libraries. As sequencing capacities continually increase, automation of library construction is becoming indispensible. We will demonstrate the automation systems IntegenX Apollo 324 (small – medium throughput) and Caliper Sciclone NGS G3 (medium – high throughput). The workshop will maintain a highly interactive environment to maximize communication among attendees as well as with lab instructors. The workshop will conclude with an evaluation of libraries and a discussion session focusing on each participant’s experimental design to help maximize efficiency and output of sequencing experiments. Course materials and lunches will be provided. To apply contact Dr. Ryan Kim (rwkim@ucdavis.edu).
