کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1163734 1490946 2015 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Multiplexed optical coding nanobeads and their application in single-molecule counting analysis for multiple gene expression analysis
ترجمه فارسی عنوان
نانوذرات کدگذاری نوری چندگانه شده و کاربرد آن در تجزیه و تحلیل چندگانه ژن در تجزیه و تحلیل شمارش تک مولکول
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• Multiplexed optical DNA coding nanobeads (MOCNBs) are fabricated.
• The MOCNBs can serve as labels for single-molecule counting analysis (SMCA).
• The limit of detection of the SMCA using MOCNBs is 1.3 × 10−15 mol L−1.
• The SMCA using MOCNBs can be used in multi-gene expression analysis.

A method for fabrication of multiplexed optical coding nanobeads (MOCNBs) was developed by hybridizing three types of coding DNAs labeled with different dyes (Cy5, FAM and AMCA) at precisely controlled ratios with biotinylated reporter DNA modified to magnetic streptavidin-coated nanobeads with a diameter of 300 nm. The color of the MOCNBs could be observed by overlapping three single-primary-color fluorescence images of the MOCNBs corresponding to emission of Cy5 (red), FAM (green) and AMCA (blue). The MOCNBs could be easily identified under a conventional fluorescence microscope. The MOCNBs with different colors could serve as the multiplexed optical coding labels for single-molecule counting analysis (SMCA) and be used in multi-gene expression analysis (MGEA). In the SMCA-based MGEA technique, multiple messenger RNAs (mRNAs) in cells could be simultaneously quantified through their complementary DNAs (cDNAs) by counting the bright dots with the same color corresponding to the single cDNA molecules labeled with the MOCNBs. We measured expression profiles of three genes from Lepidoptera insect Helicoverpa armigera in ∼100 HaEpi cells with and without steroid hormone inductions to demonstrate the SMCA-based MGEA technique using MOCNBs.

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytica Chimica Acta - Volume 886, 30 July 2015, Pages 123–132
نویسندگان
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