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author:

Ning, Jinfeng (Ning, Jinfeng.) [1] | Ye, Junlan (Ye, Junlan.) [2] | Wang, Qingqing (Wang, Qingqing.) [3] | Wang, Wei (Wang, Wei.) [4] (Scholars:王伟)

Indexed by:

EI Scopus SCIE

Abstract:

To determine multiple microRNAs (miRNAs) from cells simultaneously is essential for understanding biological functions. Capillary electrophoresis (CE) can simultaneously determine multiple miRNAs by separation. Nevertheless, similar lengths and low concentrations in cells make miRNAs hard to separate and detect. In this study, CE with laser-induced fluorescence detection was combined with catalytic hairpin assembly (CHA) to determine three miRNAs, miR-21, miR-31, and miR-122. The amplification products of CHA, which were DNA duplexes, were designed to have different lengths for different miRNAs. This allowed for easy separation of the duplexes of different miRNAs by CE. The indirect determination of miRNAs was then achieved by separating and detecting these duplexes. A magnetic field was first applied on the capillary sieving electrophoresis to assist in the separation of the duplexes. Under the optimal conditions, the three duplexes could be completely separated within 2.5 min with the detection limits of miRNAs in the range 1.12-4.05 x 10-15 M. MiR-21 and miR-31 were successfully determined from Hela cells, while miR-122 was determined from chicken livers by this method. The recoveries ranged from 97.5% to 118%. The developed method was sensitive and reliable for miRNA determination.

Keyword:

capillary electrophoresis catalytic hairpin assembly indirect determination magnetic field assistance microRNA

Community:

  • [ 1 ] [Ning, Jinfeng]Fuzhou Univ, Coll Chem, Key Lab Analyt Sci Food Safety & Biol, Minist Educ, Fuzhou, Peoples R China
  • [ 2 ] [Ning, Jinfeng]Fuzhou Univ, Coll Chem, Fujian Prov Key Lab Anal & Detect Technol Food Saf, Fuzhou, Peoples R China
  • [ 3 ] [Ye, Junlan]Fuzhou Univ, Coll Chem, Key Lab Analyt Sci Food Safety & Biol, Minist Educ, Fuzhou, Peoples R China
  • [ 4 ] [Ye, Junlan]Fuzhou Univ, Coll Chem, Fujian Prov Key Lab Anal & Detect Technol Food Saf, Fuzhou, Peoples R China
  • [ 5 ] [Wang, Qingqing]Fuzhou Univ, Coll Chem, Key Lab Analyt Sci Food Safety & Biol, Minist Educ, Fuzhou, Peoples R China
  • [ 6 ] [Wang, Qingqing]Fuzhou Univ, Coll Chem, Fujian Prov Key Lab Anal & Detect Technol Food Saf, Fuzhou, Peoples R China

Reprint 's Address:

  • [Wang, Wei]Fuzhou Univ, Coll Chem, Fuzhou 350116, Peoples R China;;

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Source :

JOURNAL OF SEPARATION SCIENCE

ISSN: 1615-9306

Year: 2024

Issue: 14

Volume: 47

2 . 8 0 0

JCR@2023

Cited Count:

WoS CC Cited Count:

SCOPUS Cited Count:

ESI Highly Cited Papers on the List: 0 Unfold All

WanFang Cited Count:

Chinese Cited Count:

30 Days PV: 2

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