Figure 6. Poly(A) tail reduction of specific maternal mRNA in aged eggs.

Poly(A) tail behavior of the indicated transcripts decreased (A) and not changed (B) upon egg aging are shown. Total mRNA from fresh (0 h) and aged (3 h) eggs was assayed by the RNA ligation-mediated poly(A) test (RL-PAT). * indicates RNaseH/oligo(dT)₂₀ digestion prior to ligation. Control lanes: –Lig, Ligation reaction performed without RNA; -RT, ligated RNA was not reverse transcribed prior to PCR. M, DNA size marker are given in base pairs. Direct sequencing of atp5a1 and tpi1 (A lower panel) reveals the actual transcript 3′ending (indicated by arrows), which is in fresh eggs at the end of the poly(A) tail (italic As), but in aged eggs several nucleotides upstream of the former end of the RNA body (clear box). P1 is the ligated primer.