Library preparation is the first step of next generation sequencing. It allows DNA or cDNA to adhere to the sequencing flow cell 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, 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, accelerating advances in research and revolutionizing our understanding of human health and disease.
Before DNA or cDNA (synthesized from RNA) can be sequenced, they must be fragmented, end-repaired, and made into sequencing libraries. Put simply, sequencing libraries are pools of DNA fragments containing adapter sequences compatible with a specific sequencing platform and indexing barcodes for individual sample identificaiton. The main library preparation methods are ligation-based library preparation tagmentation-based library preparation, and amplicon library preparation. The specific protocol you choose depends on your sequencing platform and downstream analysis. The basic steps of library preparation are fragmentation and end repair, addition of adapters, and (optional), PCR amplification:
There are several downstream applications for NGS. Choosing the appropriate library preparation protocol can significantly impact the quality and accuracy of your results. Downstream applications include:
This detailed overview walks you through major advances in sequencing technology, types of next generation sequencing, their applications and more.
Are you working in or getting started on one of these applications? See how you can easily improve your workflow and results with IDTs suite of library preparation products
Whether your research focuses on decoding the genomic sequences of complex microbial communities or identifying inherited germline single nucleotide polymorphism (SNPs) from degraded samples, xGen DNA Library Preparation Kits have simple workflows that provide confident results.
The xGen Methyl-Seq Library Prep Kit utilizes Adaptase™ technology for capturing bisulfite-converted ssDNA molecules in an unbiased manner within epigenetic research studies. The resultant libraries represent a sample base composition of the genome.
Research on differential gene expression, novel fusion genes, or alternative splicing patterns involves decoding cellular RNA. The IDT xGen NGS RNA Library Preparation Kits provide high-quality data and fast, comprehensive workflows to help you make the next discovery.
Adapters and indexing primers are key components of NGS library preparation workflows. Whether your project requires a basic indexing solution or a more sophisticated design, we have the products and expertise to deliver the right solution.