Tn5 transposase and tagmentation procedures for massively scaled sequencing projects.

ABSTRACT: Massively parallel DNA sequencing of thousands of samples in a single machine-run is now possible, but the preparation of the individual sequencing libraries is expensive and time-consuming. Tagmentation-based library construction, using the Tn5 transposase, is efficient for generating sequencing libraries but currently relies on undisclosed reagents, which severely limits development of novel applications and the execution of large-scale projects. Here, we present simple and robust procedures for Tn5 transposase production and optimized reaction conditions for tagmentation-based sequencing library construction. We further show how molecular crowding agents both modulate library lengths and enable efficient tagmentation from subpicogram amounts of cDNA. The comparison of single-cell RNA-se

SUBMITTER: Picelli S 

PROVIDER: S-EPMC4248319 | biostudies-literature | 2014 Dec

REPOSITORIES: biostudies-literature