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We have used to the kit to purify in vitro transcribed small RNAs with great success, but have not done so with purified total RNA. Here are the steps we would recommend trying with purified total RNA:1. Prepare a lysis mix with 0.7 vol. Small RNA Lysis Buffer (M1070) ａnd 0.3 vol. Binding Solution (L8042).2. Add 500 ul of lysis mix with 50 ul total RNA ａnd mix thoroughly.3. Spin at 14000 rpm for 5 min to precipitate large RNA.5. Transfer the supernatant to a new tube.6. Add 610 ul (1.1 vol.) 100% ethanol to the supernatant ａnd mix well.7.Transfer the mixture to a binding column ａnd spin 1 min to bind. Repeat the binding step with the remaining mixture.8. Wash the column first with 700 ul 100% ethanol, ａnd then with ethanol-diluted wash Solution 2.9. Dry the column ａnd elute small RNA.
We have not tested mirPremier™ microRNA Isolation Kit with sperm cells.
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