As described previously 36 (link), total RNAs from serum, plasma and blood cells of human blood samples, cell lines and cell culture medium were extracted by using a magnetic beads-based assay (MagMAX™ mirVana™ Total RNA Isolation Kit, Applied Biosystems). An appropriate amount of synthesized cel-miR-39 (RuiBiotech) was added as external standard to each sample for estimation of extraction recovery. Reverse transcription reactions were conducted with specific miRNA/piRNA stem-loop primers using the Revert Aid First Strand cDNA Synthesis Kit (Thermo).
Mai D., Zheng Y., Guo H., Ding P., Bai R., Li M., Ye Y., Zhang J., Huang X., Liu D., Sui Q., Pan L., Su J., Deng J., Wu G., Li R., Deng S., Bai Y., Ligu Y., Tan W., Wu C., Wu T., Zheng J, & Lin D. (2020). Serum piRNA-54265 is a New Biomarker for early detection and clinical surveillance of Human Colorectal Cancer. Theranostics, 10(19), 8468-8478.
Other organizations :
Sun Yat-sen University, Sun Yat-sen University Cancer Center, Huazhong University of Science and Technology, Ministry of Ecology and Environment, Chinese Academy of Medical Sciences & Peking Union Medical College
Type of sample (serum, plasma, blood cells, cell lines, cell culture medium)
dependent variables
Extracted total RNA
control variables
Amount of synthesized cel-miR-39 added as external standard for extraction recovery estimation
controls
Positive control: Synthesized cel-miR-39 as external standard
Negative control: Not specified
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