260
- RNase-free water.
- 100% ethanol.
- Centrifuges.
- RNase-free, sterile-filtered water.
- miRNA isolation kit.
- 2× hybridization buffer:100 mM 2-(N-morpholino) ethane-
sulfonic acid, 1 M [NaCl], 20 mM ethylenediaminetetraacetic
acid (EDTA), 0.01% Tween-20. Do not autoclave. Store at
2–8 °C, and protect from light. Discard solution if turned to
yellow. - 20× salt buffer: 3 M sodium chloride, 300 mM tri-sodium
citrate. Adjust pH to 7.0 with HCl. - 10% detergent solution.
- miRNA microarray slides.
- microRNA array labeling kit.
- Hybridization chamber kit.
- Hybridization gasket slide kit.
- Hybridization oven with rotation.
- Microarray scanner.
- 99% ethanol.
- Microcentrifuge.
- Vortex.
- Pipettes (positive-displacement, air-displacement, or multichan-
nel): 1–20 μL range, 20–200 μL range, 100–1000 μL range. - Pipette tips, aerosol resistant, nuclease-free: 1–20 μL range,
20–200 μL ranges, 100–1000 μL range. - RNase-free microcentrifuge tube.
- RNase-free, sterile-filtered water.
- MiRNA isolation kit.
- 20× miRNA Assay Mix.
- 10× reverse transcription buffer: 0.75 M KCl, 0.1 M MgCl 2 ,
0.5 M Tris–HCl. Adjust pH to 8.6 at 25 °C. - dNTP mix w/dTTP (100 mM total).
- RNase Inhibitor (20 U/μL).
- Reverse transcriptase.
- miRNA reverse transcription primer.
- Universal PCR Master Mix.
- Real-time thermal cycler.
2.2 Microarrays
2.3 RT-qPCR
Moein Amin et al.