Total RNA was recovered from yeast grown under 4 different conditions: YPD (to 1 OD), YPGal (to 1.7 OD), YPD (to 10 OD), and Sporulation medium.  The former two conditions show expected yields from poly(A) purification (Qiagen Oligotex kits) while the latter two consistently fail to yield RNA.  Ligation mediated poly(A) tail length analysis* shows that the distribution of poly(A) tail lengths under the first two conditions tightens from 20-120 bps to 20-60 bps in the starvation conditions (image).  To show that poly(A) length has no effect on yield from poly(A) purification, pdA20, pdA40, pdA60, and pdA80 were pooled, and 2 ug was poly(A) purified (70% yield).  Equal amounts of untreated pool and poly(A)-purified pool were separated on a denaturing 15% polyacrylamide gel and stained with ethidium bromide (image).  Quantitating the intensity of each band showed both lanes have the same distribution of each of the four oligos.
 

*LM-PAT
Denaturation:
Add Total RNA (30 ug)
Add dT12-18 (2 ug)
Heat 65 degrees for 10 min.
Ligation:
Add ligase and ligase buffer
Heat 42 degrees for 60 min.
Inactivate ligase:
Heat 90 degrees for 5 min.
Degrade RNA:
Add RNase A, RNase T1, and RNase H
Heat 37 degrees for 30 min.