Quantitative assessment of LASSO probe assembly and long-read multiplexed cloning.

ABSTRACT:

Background

Long Adapter Single-Stranded Oligonucleotide (LASSO) probes were developed as a novel tool for massively parallel cloning of kilobase-long genomic DNA sequences. LASSO dramatically improves the capture length limit of current DNA padlock probe technology from approximately 150 bps to several kbps. High-throughput LASSO capture involves the parallel assembly of thousands of probes. However, malformed probes are indiscernible from properly formed probes using gel electrophoretic techniques. Therefore, we used next-generation sequencing (NGS) to assess the efficiency of LASSO probe assembly and how it relates to the nature of DNA capture and amplification. Additionally, we introduce a simplified single target LASSO protocol using classic molecular biology techniques for qua

SUBMITTER: Shukor S 

PROVIDER: S-EPMC6657055 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature