.png){kind=icon}
Return
A single molecule array for digital targeted molecular analyses
.png){kind=icon}
OA
AI
DOI：10.1093/nar/gkn921.png)
.png){kind=icon}
.png){kind=icon}
Share.png){kind=icon}
Save
Abstract
En 中文
We present a new random array format together with a decoding scheme for targeted multiplex digital molecular analyses. DNA samples are analyzed using multiplex sets of padlock or selector probes that create circular DNA molecules upon target recognition. The circularized DNA molecules are amplified through rolling-circle amplification (RCA) to generate amplified single molecules (ASMs). A random array is generated by immobilizing all ASMs on a microscopy glass slide. The ASMs are identified and counted through serial hybridizations of small sets of tag probes, according to a combinatorial decoding scheme. We show that random array format permits at least 10 iterations of hybridization, imaging and dehybridization, a process required for the combinatorial decoding scheme. We further investigated the quantitative dynamic range and precision of the random array format. Finally, as a demonstration, the decoding scheme was applied for multiplex quantitative analysis of genomic loci in samples having verified copy-number variations. Of 31 analyzed loci, all but one were correctly identified and responded according to the known copy-number variations. The decoding strategy is generic in that the target can be any biomolecule which has been encoded into a DNA circle via a molecular probing reaction.
Keywords:
COMPARATIVE GENOMIC HYBRIDIZATION
ROLLING-CIRCLE AMPLIFICATION
PROXIMITY LIGATION
PADLOCK PROBES
IN-SITU
MULTIPLEX AMPLIFICATION
PROTEIN-DETECTION
INVERSION PROBES
DNA-MOLECULES
MICROARRAYS
AI Summary
Key information extracted from the uploaded paper, including a brief overview, abstract, background, key highlights, visual analysis, and future outlook.
Journal
IF:
13.1
Papers:
3.6W
Citations:
29.0W
Organization
Cited Papers
ShareSaveShareSaveMultiplexed proximity ligation assays to profile putative plasma biomarkers relevant to pancreatic and ovarian cancer.png)
CLINICAL CHEMISTRYOAAI
CLINICAL CHEMISTRYIF6.3
OAAIShareSaveShareSaveShareSaveHighly multiplexed molecular inversion probe genotyping: Over 10,000 targeted SNPs genotyped in a single tube assayGENOME RESEARCHOAAI
GENOME RESEARCHIF5.5
OAAIShareSavePieceMaker: selection of DNA fragments for selector-guided multiplex amplificationNUCLEIC ACIDS RESEARCHOAAI
NUCLEIC ACIDS RESEARCHIF13.1
OAAIShareSaveShareSave
View more