Next-generation sequencing (“NGS”) has revolutionized research in many areas of molecular biology, genetics, and medicine. As NGS technology has become more affordable and more widely available over the past few years, there has been increasing focus on the need for more efficient and reproducible sample preparation methods for NGS library generation. Conventional methods involve many cycles of enzymatic modification followed by purification, an arrangement that is laborious, time-consuming, and prone to template loss.
Conventional processing and purification methods in molecular biology involve nucleic acids undergoing sequential cycles of treatment followed by purification, wherein treatment and purifications are usually carried out in a separate tubes or vessels, and the overall workflow involves repeated liquid transfers (by manual or robotic pipetting devices) between the different reaction vessels. In conventional workflows, each purification step typically involves removal of the nucleic acids from the previous reaction mixture by chemical extraction, precipitation, and/or adsorption to solid phases (such as microparticles or filters). Because of the inefficiencies in the multiple liquid transfer and purification steps, poor sample yield and loss of samples due to user error are major problems for complex molecular biology workflows (like those used in NGS).