Archiving of clinical materials as Formalin-Fixed, Paraffin-Embedded (FFPE) samples is a common practice. However, the methods used for fixation and storage significantly damage the nucleic acids from these samples. Additionally, FFPE DNA samples are often available in only very small amounts. As a result, it can be challenging to construct libraries of sufficient quality and quantity to obtain good sequence data.The NEBNext product line addresses these issues in two ways:
- NEBNext® FFPE DNA Repair Mix (M6630S/L) is a cocktail of enzymes formulated to repair DNA without alteration of the DNA sequence, and is specifically optimized and validated for repair of FFPE DNA samples prior to NGS library prep.
- NEBNext Ultra™ II (E7645S/L) can be used to produce high quality libraries from low nanogram amounts of FFPE DNA samples, and is recommended for library prep when input DNA quantity and quality is limited.
This video walks you through DNA Library Preparation using the NEBNext Ultra II DNA Library Prep Kit. The video also includes tips for optimization as well as safe stopping points.
These 12 quick tips for NGS library preparation are a great refresher if you're a seasoned pro, or a great introduction if you're a beginner.