Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling
Site-specific fluorescent labeling of RNA is crucial for obtaining the structural and dynamic information of RNAs by fluorescence techniques. Post-synthetic modification of RNA based on N-hydroxysuccinimide (NHS) coupling reaction is an economic, efficient and simple strategy to introduce fluorophor...
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Online Access: | http://dx.doi.org/10.1080/21655979.2020.1765487 |
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doaj-cb319b556dc84f72bcf05cd7c1fa30ee2021-06-02T08:43:40ZengTaylor & Francis GroupBioengineered2165-59792165-59872020-01-0111159960610.1080/21655979.2020.17654871765487Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labelingMengyang Li0Shanghai Jiao Tong UniversitySite-specific fluorescent labeling of RNA is crucial for obtaining the structural and dynamic information of RNAs by fluorescence techniques. Post-synthetic modification of RNA based on N-hydroxysuccinimide (NHS) coupling reaction is an economic, efficient and simple strategy to introduce fluorophore to samples. However, this strategy are not that frequently used in RNA molecules, and the reported reaction conditions and yields varied among different systems. This study results mainly focused on screening the reaction conditions (reactants concentrations, dimethylsulfoxide concentration, solution conditions, pH and reaction time) between NHS-linked fluorophore and aminoallyl-RNA (aa-RNA) to optimize the yield of fluorescent RNA up to 55%, doubled the initial yield. What’s more, as low as one tenth of fluorescent reagent was used in our protocol compared with the reported protocols, greatly reducing the experimental cost. The protocol can be applied as a general guide potentially for RNA labeling by NHS-ester coupling reaction.http://dx.doi.org/10.1080/21655979.2020.1765487fluorescent labelingn-hydroxysuccinimidenhs-linked fluorophorerna labeling |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Mengyang Li |
spellingShingle |
Mengyang Li Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling Bioengineered fluorescent labeling n-hydroxysuccinimide nhs-linked fluorophore rna labeling |
author_facet |
Mengyang Li |
author_sort |
Mengyang Li |
title |
Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling |
title_short |
Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling |
title_full |
Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling |
title_fullStr |
Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling |
title_full_unstemmed |
Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling |
title_sort |
optimization of n-hydroxysuccinimide ester coupling with aminoallyl-modified rna for fluorescent labeling |
publisher |
Taylor & Francis Group |
series |
Bioengineered |
issn |
2165-5979 2165-5987 |
publishDate |
2020-01-01 |
description |
Site-specific fluorescent labeling of RNA is crucial for obtaining the structural and dynamic information of RNAs by fluorescence techniques. Post-synthetic modification of RNA based on N-hydroxysuccinimide (NHS) coupling reaction is an economic, efficient and simple strategy to introduce fluorophore to samples. However, this strategy are not that frequently used in RNA molecules, and the reported reaction conditions and yields varied among different systems. This study results mainly focused on screening the reaction conditions (reactants concentrations, dimethylsulfoxide concentration, solution conditions, pH and reaction time) between NHS-linked fluorophore and aminoallyl-RNA (aa-RNA) to optimize the yield of fluorescent RNA up to 55%, doubled the initial yield. What’s more, as low as one tenth of fluorescent reagent was used in our protocol compared with the reported protocols, greatly reducing the experimental cost. The protocol can be applied as a general guide potentially for RNA labeling by NHS-ester coupling reaction. |
topic |
fluorescent labeling n-hydroxysuccinimide nhs-linked fluorophore rna labeling |
url |
http://dx.doi.org/10.1080/21655979.2020.1765487 |
work_keys_str_mv |
AT mengyangli optimizationofnhydroxysuccinimideestercouplingwithaminoallylmodifiedrnaforfluorescentlabeling |
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1721406278453952512 |