NGS Library Preparation: Theory and Practice

Previously recorded on February 17,2015. Presented by Dr. Maryke Appel of Kapa Biosystems.

Part 1: The Objectives of Library Construction

Optimized Library Construction for High-quality Transcriptome Sequencing on the Illumina® Platform

Previously recorded on January 22, 2015. Presented by Dr. Adriana Geldart of Kapa Biosystems. High-quality library preparation is key in transcriptome sequencing to ensure that the data acquired at the end of sequencing is accurate. Dr. Geldart presents data on the ideal quality control metrics to use when preparing RNA-Seq libraries, and show that these metrics assist in predicting the outcome of the sequencing run. We would like to thank Dr. Tom Westerling for his collaboration in this webinar. Products discussed in this presentation:

Increasing percentage of analyzed patients using a highly streamlined library prep workflow for exome and long-insert libraries

Previously recorded on December 17, 2014. Presented by Dr. Jonathan J. Keats of the Translational Genomics Research Institute. Multiple myeloma is a malignancy characterized by the accumulation of plasma cells in the bone marrow. This tumor expansion results in a suppressed white blood cell system, anemia, severe osteoporosis, and compromises kidney function. Recent advances in next generation sequencing can now identify nearly all genetics events existing in an individual case of multiple myeloma. Products discussed in this presentation:

FFPE library construction for Illumina® sequencing: insights and improvements

Previously recorded on May 1, 2014. Formalin-fixed paraffin embedded (FFPE) tissue is an important source of DNA for cancer genomics studies and clinical diagnostics. One of the major challenges of high-throughput sequencing is the ability to process low-input samples of variable quality with predictable success rates in standard sample preparation pipelines. Products discussed in this presentation:

Meeting the Challenges of High-throughput, Low-input Library Construction for Illumina Sequencing

Part 1: (18 minutes)

Dr. Appel discusses Evolved Enzymes and Optimized Protocols to achieve high library diversity and low-bias library amplification, while meeting challenges such as:

  • Maximizing yield of adapter-ligated libraries
  • Minimizing the number of library amplification cycles
  • Reducing amplification bias, PCR duplication rates, substitutions and other amplification artifacts

Part 2: (16 minutes)

Dr. Appel discusses building a protocol that is scalable over a wide range of DNA inputs and also automation friendly. She introduces case studies of four liquid handling platforms:

  • Caliper Sciclone NGS Workstation
  • Beckman Coulter Biomek FXP
  • Agilent Bravo B NGS Workstation
  • Eppendorf epMotion 5075 TMX