The basic steps of a NGS library are as follows" 1.Sample extraction 2. DNA Fragmentation 3. End Repair and Adapter Ligation 4. Size Selection (varies according to different platforms chosen) 5. Library Amplification (by PCR) 6. Library QC (by qPCR, fluorometry, spectrophotometry and gel electrophoresis) 7. Library Pooling/ Multiplexing 8. Sequencing
Of course there are various cleanups and QC s being performed at different steps as necessary. At what step do we decide for depth and coverage (depends on applications such as whole genome or exome etc.),? Is there any calculation for Library pooling? Does above cover general library steps or is something missing?