Before you start
Next generation sequencing is based on genetic material. Before you can prepare libraries, you must have isolated and purified genetic material. Check your DNA or RNA extraction kit for compatibility with NGS before extraction. Extraction and purification
must result in adequate yield and quality for successful sequencing. For RNA sequencing, RNA must first be converted to cDNA.
Step 1: Library prep
The library preparation step allows your samples to be processed on your sequencer.
IDT products are compatible with many types of sequencing platforms, including Illumina, Pacific Biosciences, and Oxford Nanopore. The library prep step fragments the sample into shorter sequences to combat sequencing bias and attaches adapters to make the sequence compatible with the sequencer. Identifying sequences called “barcodes” or “indexes” can also be added to the
samples. Barcodes allow the combining of samples, also called “pooling” or “multiplexing,” so that they can be sequenced simultaneously, saving time and money.
An alternative to whole genome sequencing (WGS) is
targeted sequencing. Instead of sequencing the entire genome
of your sample, you can choose specific portions of the genome to focus your study for a faster, more efficient experiment. Preparing libraries for targeted sequencing is called enrichment. Enrichment can occur either during library prep (amplicon sequencing) or after library prep (hybridization capture), depending on the enrichment method chosen.