19. Phenol:chloroform:isoamyl alcohol (25:24:1), pH 4.3.
20. Ethanol.
21. 3 M NaAc prepared with RNase-free water.
22. Glycogen.
23. 1Â TBE buffer.
24. FA buffer.
25. Urea-polyacrylamide gel reagents.
26. 0.4 M NaCl prepared with RNase-free water.
27. RNA dissolve buffer: 100 mM KCl, 30 mM Tris–HCl, pH 7.5
in RNase-free water.
2.4 RNA Extraction
1. Mortar and pestle.
2. Lab spoons.
3. Scale.
4. Liquid nitrogen.
5. Heat blocks.
6. Centrifuge.
7. Gel electrophoresis casts, trays, combs, tank, and power device.
8. 1.5 mL Eppendorf tubes and all size tips.
9. Vortex.
10. Trizol.
11. Phenol:chloroform:isoamyl alcohol (25:24:1), pH 4.3.
12. Chloroform.
13. Ethanol.
14. RNase-free water.
15. Autoclaved double-distilled water.
16. 75% Ethanol prepared with RNase-free water.
17. FA buffer.
18. Agarose.
19. Ethidium bromide (EB).
2.5 Preparation of
RNA with Radiolabeled
Marker Cocktail and
Size Selection
1. The same materials as described in Subheading 2.3.2, except
T4 PNK and [γ32
P] ATP.
2. Labeled markers described in Subheading 3.3.2.
3. RNA samples as described in Subheading 3.4.
2.6 3
0 Ligation
1. The same materials as described in Subheading 2.3.2, except
T4 PNK and [γ32
P] ATP.
2. 3
0 Adapter synthesized described in Subheading 3.1.
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