Library Preparation for Illumina

Illumina^® sequencing instruments can be found in laboratories of all sizes, from lab-scale to production-scale, and NEBNext kits can be used to construct the high-quality libraries required for them all, for a wide array of sample types and applications. 

NGS Library Preperation

NGS library preparation for the Illumina^® sequencing platform requires inputs of a defined length and, therefore, fragmentation of DNA, the use of cDNA prepared from RNA, or targeted amplicons are the starting point. This is followed by end repair (3´ and 5´) to generate blunt-ended, phosphorylated molecules, followed by the addition of a non-templated d(A)-tail before ligation to an adaptor. Depending on the amount of available sample and the experimental goals, the final step may include PCR amplification of the library; when sufficient amounts of library are available without PCR, PCR-free workflows can be employed. Small RNA libraries are constructed using a different workflow, in which adaptors are ligated directly to the small RNA molecules, followed by reverse transcription and PCR amplification.  
 
The NEBNext^® suite of products supports Illumina sequencing with sample preparation tools that streamline workflows, minimize inputs, and improve library yields and quality. NEBNext sample preparation kits are available for both DNA library prep and RNA library prep; kits and modules are available for genomic DNA, FFPE DNA, methylated DNA, microbiome DNA, total RNA, single cell/low input RNA, and small RNA samples. These are complemented by a wide range of index primers and index adaptors for multiplexing. In addition to the extensive QCs performed on individual kit components, all NEBNext kits for Illumina library prep are functionally validated by preparation of a library, followed by Illumina sequencing. NEBNext library preparation reagents are available in multiple kit and workflow formats, for maximum convenience and flexibility. Designed for use in concert with NEBNext Multiplex Oligos for simple yet powerful NGS sample multiplexing. 
 

NEBNext Ultra™ II is at the heart of our DNA and RNA library prep workflows.

The NEBNext Ultra II reagents and workflow are at the heart of NEB’s portfolio for next generation sequencing library preparation. NEBNext Ultra II kits and modules for Illumina^® are an optimized combination of reagents and simplified workflows, enabling you to create DNA or RNA libraries of highest quality and yield, even when starting from extremely low or low-quality input amounts. 

The Ultra II workflow is at the heart of:  

• Ultra II DNA & FS DNA Library Prep, with PCR-free workflow options
• Enzymatic Methyl-seq
• Ultra II RNA & Directional RNA Library Prep
• Single Cell/Low Input RNA Library Prep  
• NEBNext ARTIC SARS-CoV-2 library prep 
• Modular options for each step in the workflow
  
  

Don’t just take our word for it; review the nearly 20,000 publications featuring NEBNext reagents, instead.