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MicroRNA isolation kit (A&A Biotechnology) following the manufacturers’ protocol |
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Protocol tips |
MicroRNA isolation kit (A&A Biotechnology) following the manufacturers’ protocol |
Publication protocol
MicroRNA isolation kit requires a phenol/chloroform extraction step and purification is based on silica matrix columns. 3 × 107 cells from mid‐log culture were pelleted and treated with 800 μL of Fenozol. The lysed cells were incubated at 50 °C for 5 min. 200 μL of chloroform was added and left at the room temperature for 3 min. The probes were centrifuged for 10 min at 10 000 g and nucleic acids within the resulting supernatant were precipitated with 1/3 volume of isopropanol. This mixture was filtered through the silica columns by centrifugation for 1 min at 10 000 g. High molecular weight (HMW) RNA remained bound to the columns. Low molecular weight (LMW) RNA from the flow through were precipitated with 2/3 volume of isopropanol and filtered through the silica columns by centrifugation for 1 min at 10 000 g. LMW RNA were recovered from the columns with 50 μL of DEPC‐treated water.
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Papers
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Manufacturer protocol
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