10. 0.2 M Ethylenediaminetetraacetic acid (EDTA), pH 8.0.
11. 100% Ethanol.
12. 3 M Sodium acetate (pH 5.5).
13. 75% Ethanol.
14. NanoDrop™ 2000/2000c Spectrophotometer.
15. PCR thermocycler.
16. 0.2 mL PCR tubes.
17. 1.5 mL Microcentrifuge tubes.
18. Benchtop centrifuge capable of speeds up to 13,000 Â g.
19. Agarose.
20. 1Â Tris-acetate-EDTA (TAE) buffer: 40 mM Tris-acetate and
1 mM EDTA.
21. 6Â Loading dye.
22. 1 kb DNA ladder.
23. 10 mg/mL Ethidium bromide.
24. Erlenmeyer flask.
25. Microwave.
26. Equipment and supplies for running an agarose gel including
gel casting mold, well combs, and horizontal electrophoresis
chamber.
27. Voltage source.
28. UV light source.
29. À20
C Freezer.
30. 4
C Refrigerator.
31. Incubator set to 37
C.
2.2 Preparation
of Fluorescent sRNAs
1. ShortCut
® RNase III.
2. 10Â ShortCut reaction buffer.
3. 200 mM MnCl 2 .
4. 250 mM Ethylenediaminetetraacetic acid (EDTA).
5. 100% Ethanol.
6. 80% Ethanol.
7. Nuclease-free water.
8. NanoDrop™ 2000/2000c Spectrophotometer.
9. Incubator set to 37
C.
10. 40% Acrylamide solution for polyacrylamide gel electrophoresis (PAGE).
11. 10Â TBE buffer: 890 mM Tris base, 30 mM EDTA, pH 8.0,
890 mM boric acid.
Visualizing Fungal RNA Uptake
217