DNA for Illumina®

Library preparation for the Illumina® sequencing platform requires fragmentation of DNA, repair of 3´ and 5´ ends to form blunt-ended, phosphorylated molecules, and the addition of a non-templated dA-tail before ligation to an adaptor. If necessary to achieve sufficient yields, the final step is PCR amplification of the library. For more detail, select a workflow tab below.

NEBNext^® kits are available for sample preparation of genomic DNA, ChIP DNA, methylated DNA, microbiome DNA, and FFPE DNA. These are complemented by a wide range of index primers, for multiplexing. Setting a new standard, our NEBNext Ultra™ II FS DNA kits include a new fragmentation reagent, which is combined with end-repair and dA-tailing reagents, enabling these steps to be performed in the same tube, without any clean-ups (and therefore without any sample loss). The same fragmentation protocol is used for any input amount from 100 pg to 500 ng, and DNA of any GC content for added ease. The high-efficiency Ultra II ligation reagents and Q5® Ultra II PCR master mix complete the library prep workflow, resulting in a fast, high-yield, high-quality library prep workflow that is completely scalable. 

NEBNext Enzymatic Methyl-seq, an enzyme-based alternative to bisulfite sequencing employs Ultra II reagents and enables superior detection of 5mC and 5hmC, from fewer sequencing reads. Details on this workflow can be found below.