Materials

1. Universal extraction bags 12 x 14 cm with synthetic intermediate layer (Bioreba AG).

2. Dulbecco's phosphate-buffered saline without magnesium or calcium (PBS; pH 7.4): 8 g of NaCl, 0.15 g of KH2PO4, 2.9 g of Na2HPO412H2O, 0.2 g of KCl per liter.

3. Sample extraction buffer (SEB): PBS (pH 7.4) containing 0.05% Tween-20, 2% (w/v) polyvinylpyrrolidone 25000, 0.2% (w/v) ovalbumin, and 0.5% (w/v) bovine serum albumin.

4. HOMEX 6 Plant Tissue Homogenizer (Bioreba AG).

6. Coating buffer (pH 9.6): 0.16% (w/v) Na2CO3, 0.29% (w/v) NaHCO3.

7. Antibodies at 1 mg/mL, diluted to a working concentration of 1-in 100 in coating buffer (pH 9.6).

8. Plate seals.

10. Enzyme solution: Cellulase "onozuka" RS/Macerozyme R-10 (Yakult Pharmaceuticals) 0.1 g of each enzyme in 1 mL of RNase-free water.

11. 1X RT buffer: 50 ml Tris-HCl (pH 8.3), 75 ml KCl, 3 ml MgCl2.

12. 5X RT buffer: 250 ml Tris-HCl (pH 8.3), 375 ml KCl, 15 ml MgCl2.

13. Moloney murine leukemia virus reverse transcriptase.

14. NotI poly dT primer; 5'-CAATTCGCGGCCGCT18-3'.

15. dNTPs, 10 ml each with respect to dATP, dCTP, dGTP, and dTTP.

16. RNase-free H2O.

17. 100 ml dithiothreitol (DTT).

18. Recombinant RNasin ribonuclease inhibitor (Promega).

19. 10X PCR buffer.

22. Taq DNA polymerase 5U/|L.

23. Electrophoresis equipment.

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