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Extraction and Detection of mRNA from a Single K562 Cell Based on the Functionalized Superparamagnetic Nanoparticles
Author(s) -
ZHU LongZhang,
YU PingGuo,
SHEN HeBai,
JIA NengQin,
LONG DeHong,
ZHOU HaiQing
Publication year - 2008
Publication title -
chinese journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.28
H-Index - 41
eISSN - 1614-7065
pISSN - 1001-604X
DOI - 10.1002/cjoc.200890185
Subject(s) - chemistry , superparamagnetism , lysis , messenger rna , agarose gel electrophoresis , nanoparticle , agarose , magnetite , chromatography , magnetic nanoparticles , microbiology and biotechnology , dna , nanotechnology , biochemistry , gene , materials science , magnetic field , magnetization , physics , quantum mechanics , biology , metallurgy
Abstract A novel and promising method was developed to extract mRNA from a single cell based on the functionalized superparamagnetic nanoparticles. The oligo(dT)‐coupled magnetite nanobeads were synthesized by the reaction of oligo(dT) and thiol‐modified γ ‐Fe 2 O 3 nanoparticles. The single cell was isolated from the massive cultivation according to a semi‐quantum approaching technique and then lysed before mRNA separation. The oligo(dT)‐coupled magnetite nanobeads were added to the crude lysates and then magnetic separation was preformed to get mRNA. The mRNA amplification through a two‐step RT‐PCR method was achieved. The agarose gel electrophoresis of PCR products after amplification shows that mRNA could be extracted from a single cell successfully.