miRNeasy Mini kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step

Protocol tips
- To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step

Publication protocol

Total RNA was extracted from A2780 and A2780/DDP cells using Trizol (Invitrogen, USA) and miRNeasy mini kit (QIAGEN, Denmark) according to manufacturer’s instructions. The quality and quantity were measured by the spectrophotometer (Beckman Coulter, DU730, USA) and RNA integrity was verified by gel electrophoresis. RNA (1ug) was labeled with Hy3 fluorescent using the miRCURY Power labeling kit (Exiqon, Vedbaek, Denmark) and then hybridized on the miRCURYLNA Array (v.18.0, Exiqon) containing 1887 human miRNA capture probes annotated in miRBase 18.0. Following hybridization, the fluorescent signal images were acquired using a Genepix 4000B scanner (Axon Instruments,Union City, CA, USA) and were analyzed with Genepix Pro 6.0 soft-ware (Axon Instruments), in which the median normalization was obtained. Two-fold or larger change was set as a threshold of significant difference.

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Manufacturer protocol

Download the product protocol from Qiagen for miRNeasy Mini kit below.

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