The Proceedings of Mechanical Engineering Congress, Japan
Online ISSN : 2424-2667
ISSN-L : 2424-2667
2015
Session ID : J0540101
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J0540101 Purity of cytoplasmic RNA extracted from single cells via electrical lysis and isotachophoresis
Shota HATARyuji YOKOKAWAHidetoshi KOTERAHirofumi SHINTAKU
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Abstract
We report on our efforts to measure the purity of RNA extracted from single cells via electrical lysis and ITP (isotachophoresis). We have developed a fluorescence microscopy to quantify absolute masses of RNA and protein focused at ITP interface with fluorescence dyes that specifically bind with RNA and protein, respectively. This quantification is performed without enzymatic amplification in less than 5 min. We demonstrate our technique using K562 single cells, for which we obtained an average of 12.1±1.6 pg and 43.8±8.8 pg of RNA and protein masses, respectively, and 36.3±4.9% of RNA purity.
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© 2015 The Japan Society of Mechanical Engineers
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