What is the typical library size distribution for QuantSeq FFPE libraries?
The library shape, size, and yield may vary depending on the sample input and quality. Heavily degraded FFPE samples typically generate shorter libraries than FFPE samples of higher quality.
Most inserts are expected to be >75 bp, corresponding to final libraries with fragment sizes >200 bp.
The figure below depicts examples of QuantSeq FFPE libraries obtained from various FFPE RNA samples differing in RNA input amounts and quality:
Figure | Bioanalyzer traces of QuantSeq FFPE libraries synthesized from 20 ng Homo sapiens (human, Hs) kidney FFPE RNA (RIN 2.9, DV200 50 %, red trace), 20 ng Hs brain FFPE RNA (RIN 5.5, DV200 91 %, blue trace), 10 ng Mus musculus (mouse, Mm) Liver FFPE RNA (RIN 2.3, DV200 83 %, turquoise trace) and 10 ng Mm Liver fresh frozen RNA (RIN 9.2, green trace). Due to the highly fragmented nature of FFPE RNA libraries are significantly shorter than the ones generated from fresh frozen RNA.