Figure 2
From: Naturally occurring antisense RNA of histone H2a in mouse cultured cell lines
(a) RT-PCR products from cDNAs obtained by priming total RNA with random hexamers. Lanes: 1, DNA ladder (100-bp ladder, TOYOBO); 2–7, RT-PCR products amplified with primers F1 and R1 (upper) and those amplified with primers F1 and R2 (lower). RNA was extracted from Hepa 1–6 (lanes 2 and 5), 3T3 (lanes 3 and 6), and LLC (lanes 4 and 7) cells. Superscript III was not added in the reaction of lanes 2–4. Lane 8, PCR product of genomic DNA amplified with primers F1 and R1 (upper) and that amplified with primers F1 and R2 (lower). Arrows indicate the expected products. (b) Patterns of digestion of PCR products by Pst I. Lanes: 1, DNA ladder; 2–4, digests of PCR products amplified with primers F1 and R1. PCR product was produced from Hepa 1–6 (lane 2), 3T3 (lane 3), and LLC (lane 4). Lanes 5–7, digests of PCR products amplified with primers F1 and R2. PCR product was produced from Hepa 1–6 (lane 5), 3T3 (lane 6), and LLC (lane 7). Arrows indicate the expected products. (c) RT-PCR products and the Eco RI-digest patterns of cDNAs obtained by priming total RNA with the specific primer R3. Lanes: 1 and 8, DNA ladder; 2–7, RT-PCR products amplified with primers F2 and R3. RNA was extracted from Hepa 1–6 (lanes 2 and 5), 3T3 (lanes 3 and 6), and LLC (lanes 4 and 7). Superscript III was not added in the reaction of lanes 2–4. Lanes 9–11, Eco RI-digests of PCR products amplified with primers F2 and R3. PCR product was produced from Hepa 1–6 (lane 9), 3T3 (lane 10), and LLC (lane 11). Arrows indicate the expected products.