Toehold-enhanced LNA probes for selective pull down and single-molecule analysis of native chromatin.

ABSTRACT: The organization of DNA into chromatin is thought to regulate gene expression in eukaryotes. To study its structure in vitro, there is a need for techniques that can isolate specific chromosomal loci of natively assembled chromatin. Current purification methods often involve chemical cross-linking to preserve the chromatin composition. However, such cross-linking may affect the native structure. It also impedes single molecule force spectroscopy experiments, which have been instrumental to probe chromatin folding. Here we present a method for the incorporation of affinity tags, such as biotin, into native nucleoprotein fragments based on their DNA sequence, and subsequent single molecule analysis by magnetic tweezers. DNA oligos with several Locked Nucleic Acid (LNA) nucleotides are shown

SUBMITTER: Hermans N 

PROVIDER: S-EPMC5711847 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature