NEBNext Ultra II produces the highest yield libraries from a broad range of input amountsA.
|FFPE DNA||DNA INPUT (ng)||LIBRARY YIELDS IN ng||% MAPPED||% MAPPED IN PAIRS||% DUPLICATION||% CHIMERAS|
Libraries were prepared from 17–30 ng of human DNA extracted from the FFPE tissue samples listed, amplified using 10 cycles of PCR and sequenced on the Illumina MiSeq®. This data illustrates that NEBNext Ultra II enables high quality sequence data, even with low input amounts of FFPE DNA.
A: Reads were mapped to the GRCh37 reference genome using Bowtie 2.2.4.
% Mapped: The percentage of reads mapped to Human GRCh37 reference.
% Mapped in Pairs: The percentage of reads whose mate pair was also aligned to the reference.
% Duplication: The percentage of mapped sequence that is marked as duplicate.
% Chimeras: The percentage of reads that map outside of a maximum insert size or that have the two ends mapping to different chromosomes.
B: Bioanalyzer® traces of each library show high quality libraries with minimal adaptor-dimer.
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.