Library preparation is the first step of next generation sequencing. It allows DNA or RNA to adhere to the sequencing flowcell and allows the sample to be identified. Two common methods of library preparation are ligation-based library prep and tagmentation-based library prep. Once your libraries are prepared, you will be ready for the next step in your next generation sequencing workflow.
Next generation sequencing, or high-throughput sequencing, enables sequence profiling of everything from genomes and transcriptomes to DNA-protein interactions. The technologies used are an integral part of research and discovery in biology. The ability to generate large amounts of sequence data in a relatively short amount of time enables a wide range of applications and accelerates advances in research and the clinic, which has revolutionized understanding of human health and treatment of disease.
Before DNA or RNA samples can be sequenced by next generation sequencing, they must be fragmented, end-repaired, and collected into adapter-ligated libraries. Library preparation protocols can influence the results generated by your NGS experiments. The major steps of ligation-based library preparation are summarized as follows:
There are two common methods of DNA library preparation.
Ligation: DNA fragmentation and ligation of adapters to the ends of the fragments in two separate steps.
Tagmentation: DNA fragmentation and adapter ligation are combined into one reaction step.