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The method developer's guide to oligonucleotide design

EXPERT REVIEW OF PROTEOMICS(2024)

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Abstract
IntroductionDevelopment of new methods is essential to make great leaps in science, opening up new avenues for research, but the process behind method development is seldom described.Areas coveredOver the last twenty years we have been developing several new methods, such as in situ PLA, proxHCR, and MolBoolean, using oligonucleotide-conjugated antibodies to visualize protein-protein interactions. Herein, we describe the rationale behind the oligonucleotide systems of these methods. The main objective of this paper is to provide researchers with a description on how we thought when we designed those methods. We also describe in detail how the methods work and how one should interpret results.Expert opinionUnderstanding how the methods work is important in selecting an appropriate method for your experiments. We also hope that this paper may be an inspiration for young researchers to enter the field of method development. Seeing a problem is a motivation to develop a solution.
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Key words
Method development,oligonucleotide design,microscopy,Protein-protein interactions,antibodies,proximity ligation
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