Simple, streamlined, cost-effective cDNA synthesis method from cell cultures.

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Tác giả: Nikolina Canová, Veronika Hanzíková, Markéta Kuklová, Jiří Novotný, Ladislav Šenolt, Ondřej Slanař, Monika Šteigerová, Daniel Stránský

Ngôn ngữ: eng

Ký hiệu phân loại: 782.96 +Speaking voices (Choral speech)

Thông tin xuất bản: England : Open biology , 2025

Mô tả vật lý:

Bộ sưu tập: NCBI

ID: 695709

Applications like drug development need simple and streamlined methods to process samples from 96-well cell culture plates for gene expression measurements. Unfortunately, current options are expensive for such processing. Therefore, our aim was to develop a method that would allow streamlined analysis of mRNA from 96-well cell culture plates while being relatively cheap and simple. We developed a method based on the qPCR 'Cells-to-cDNA' approach and validated it against commercially available kits using the same approach or spin columns-based RNA purification. For this purpose, we conducted a series of comparisons of gene expression from peripheral blood mononuclear cells, SK-HEP-1 and U-87 cell cultures in 96-well plates. Our final method involved lysing cells with 25-100 µl solution of 0.5% SDS, 10 mM DTT, 1 mg ml
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