Before DNA or RNA samples can be sequenced, they must be fragmented, end-repaired, and ligated to sequencing adapters. Library preparation protocols can influence the results generated by your next generation sequencing data. The major steps of ligation-based library preparation are pictured below (Figure 1) and summarized as follows:
Fragmenting shears the samples into pieces of DNA of a desired length. Typically, whole genome sequencing works best with 350 bp fragments, and hybridization capture works best with 200 bp fragments.
There are 2 main methods of fragmentation for ligation-based DNA library prep:
Tagmentation is an alternative protocol that combines the fragmentation and adapter ligation steps. Some versions of this protocol could introduce a coverage bias that, depending on experimental design, may impact sequencing results.
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