Table 1. Titration methods used for Next Generation Sequencing.

Library Titration Method	Requires Bio-analyzer analysis	Quantification Method	Sensitivity	DNA Input per Lane
QuBit	yes	Estimation through Average Library Molecular Size	pg-ng	8 pM
PicoGreen	yes	Estimation through Average Library Molecular Size	pg-ng	8 pM
qPCR	yes	Estimation through Average Library Molecular Size	0.1-1 pg	8 pM
ddPCR	no	Count of number of molecules per microliter	fM	8 pM

NGS consists of massive parallel sequencing of libraries from shRNA screen to ChIPseq, Exome -, Whole-genome sequencing and chromatin capture such as HiC and 4C. This table reports the most common methods for titration, including dsDNA quantification by spectrofluoremetry (QuBit, PicoGreen); real-time PCRs (qPCR, KapaBiosystems); and finally ddPCR (BioRad). Using the advantages of an emulsion PCR approach, ddPCR-based titration, unlike the others does not require a high sensitivity electrophoresis analysis (BioAnalyzer, Agilent) in order to determine the average molecular size of the library for calculation of molarity. After accurate quantification, 8 pM–12 pM of DNA is loaded per lane in the deep sequencer (Illumina’s Recommendations).