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靶标交替 SELEX:利用交替靶标筛选以生成针对保守末端二肽的适体。

Target-switch SELEX: Screening with alternating targets to generate aptamers to conserved terminal dipeptides.

机构信息

Google, LLC, Mountain View, CA 94043, USA.

Google, LLC, Mountain View, CA 94043, USA.

出版信息

STAR Protoc. 2022 Dec 16;3(4):101724. doi: 10.1016/j.xpro.2022.101724. Epub 2022 Oct 7.

DOI:10.1016/j.xpro.2022.101724
PMID:36208449
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9557731/
Abstract

Systematic evolution of ligands by exponential enrichment (SELEX) encompasses a wide variety of high-throughput screening techniques for producing nucleic acid binders to molecular targets through directed evolution. We describe here the design and selection steps for discovery of DNA aptamers with specificity for the two consecutive N-terminal amino acids (AAs) of a small peptide (8-10 amino acids). This bead-based method may be adapted for applications requiring binders which recognize a specific portion of the desired target. For complete details on the use and execution of this protocol, please refer to Hong et al. (2022).

摘要

系统进化的配体指数富集(SELEX)涵盖了广泛的高通量筛选技术,通过定向进化生产与分子靶标结合的核酸配体。我们在这里描述了设计和选择步骤,用于发现对小肽(8-10 个氨基酸)的两个连续 N 端氨基酸(AA)具有特异性的 DNA 适体。这种基于珠粒的方法可以适用于需要识别所需靶标特定部分的结合物的应用。有关此方案使用和执行的完整详细信息,请参阅 Hong 等人(2022 年)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/c810b6b11388/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/4079424317c6/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/e4f1b61fec2e/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/99cb7f5478ce/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/c9e148d39c72/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/e73ed97d1303/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/c13b155bdbe5/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/7d734e4709f2/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/c810b6b11388/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/4079424317c6/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/e4f1b61fec2e/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/99cb7f5478ce/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/c9e148d39c72/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/e73ed97d1303/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/c13b155bdbe5/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/7d734e4709f2/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd31/9557731/c810b6b11388/gr7.jpg

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Investigating the Influences of Random-Region Length on Aptamer Selection Efficiency Based on Capillary Electrophoresis-SELEX and High-Throughput Sequencing.
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Anal Chem. 2021 Dec 28;93(51):17030-17035. doi: 10.1021/acs.analchem.1c03661. Epub 2021 Dec 15.
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How to measure and evaluate binding affinities.如何测量和评估结合亲和力。
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