FIG. 3.

Primer extension of HSP82 mRNA from total RNA. Total RNA was isolated from exponentially growing yeast cultures at 25°C or after the culture had been shifted to 39°C for 30 min. The amount of total RNA was quantified, and equal amounts (30 μg) of RNA were used for primer extension with ³²P end-labeled primers for both ACT1 and HSP82. The left panel shows primer extension reactions with an actin-specific primer and a primer specific for the 5′ end of HSP82. Actin was used as an internal control due to its long half-life in vivo. The right panel shows a primer extension reaction with a primer specific for the 3′ end of HSP82. NHS, RNA extracted from yeast under non-heat shock conditions; HS, RNA extracted from yeast under heat shock conditions.