FFPE DNA poses many challenges for library preparation, including low input amounts and highly variable damage from fixation, storage, and extraction methods. Regions of interest are often enriched using hybrid capture-based approaches, and these workflows require a high input of diverse, uniform DNA library.
The NEBNext Ultrashear™ FFPE DNA Library Prep Kit incorporates enzymatic fragmentation of FFPE DNA using NEBNext UltraShear, a new enzyme mix optimized for use with FFPE DNA that increases library yields and quality, while improving scalability and ease of use. The kit also repairs damage to DNA caused by the FFPE process and includes reagents and a protocol optimized for library prep of FFPE DNA. The workflow is streamlined and user-friendly, with minimal hands-on time, and the kit improves yields and quality of libraries from a range of FFPE DNA sample input (5-250 ng) and quality.
View or download extensive performance data in our Data Supplement.
Includes specialized enzymatic fragmentation and FFPE DNA repair reagents, plus optimized library prep reagents and protocol
Increased library yields
Improved sequencing metrics
Greater sensitivity of somatic variant calling
5-250 ng input range
Automation-friendly workflow
For FFPE DNA library prep without enzymatic fragmentation, the NEBNext FFPE DNA Library Prep Kit (NEB #E6650) is available.
View or download extensive performance data in our Data Supplement.
FFPE DNA is often compromised in quality and quantity and is challenging for library prep and sequencing, including in target enrichment workflows that require high amounts of library input. The NEBNext UltraShear FFPE DNA Library Prep Kit addresses these challenges and improves ease of workflow and scalability:
Repairs FFPE-induced damage using the NEBNext FFPE DNA Repair Mix v2
Incorporates specialized enzymatic DNA fragmentation using NEBNext UltraShear
Uses NEBNext Ultra II library prep reagents in a protocol optimized for FFPE DNA
Robustly amplifies libraries using NEBNext MSTC™ FFPE Master Mix
High library yields, sufficient for input into target enrichment workflows
User-friendly workflow with minimal hands-on time and multiple safe stopping points
The NEBNext UltraShear FFPE DNA Library Prep Kit reduces damage from deamination and oxidation due to the fixation, storage, and extraction process for FFPE DNA samples, thereby reducing false positive variant calls.
The combination of specialized enzymatic FFPE DNA fragmentation and repair with optimized reagents and protocols provides superior performance with this challenging sample type.
For FFPE DNA library prep without enzymatic fragmentation, the NEBNext FFPE DNA Library Prep Kit (NEB #E6650) is available.
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