What are the major steps in the xGen™ DNA Library Prep Kit EZ library construction?
Enzymatic prep. Consists of fragmentation of dsDNA followed by end-repair and dA-tailing of the fragments—all performed in a single reaction.
Adapter Ligation. Stubby or full-length Y adapters are ligated to 3’-dA-tailed molecules.
Library amplification. Employs a high-fidelity, low-bias polymerase. For indexing using the ligation workflow, amplification primers can be used, but PCR is optional when using the minimum of 100 ng input. For indexing by PCR, stubby Y adapters are used during ligation and amplified with indexing primers to complete the adapter sequence and add sample indexes.