Fig 8. In situ PCR for the localization and expression studies of osa-miR12477.

Paraffin embedded leaves of the control (C) and 256 mM NaCl treated (T) Pokkali seedlings were sectioned (60 μm) on a microtome, processed to remove the paraffin, osa-miR12477 was reverse transcribed using miRNA specific primer and amplified with miRNA specific stem-loop forward primer and universal reverse primer using high fidelity Taq polymerase with dig-11-dUTP (Roche). The section was probed with anti-DIG-AP-Fab fragment and the miRNA expression was visualized using BM-purple substrate (Roche) on a light microscope. The leaf sections of both control (C) and NaCl treated (T) seedlings were processed without (A) and with (B) osa-miR12477 specific stem-loop primer.