Methodologies for In Vitro Cloning of Small RNAs and Application for Plant Genome(s)
Figure 6
Isolation and cloning of small RNAs from cotton ovule tissue libraries [33]: (a) the example of 15% denaturing PAGE electrophoresis of total RNA from developing ovules at different DPA (0 to 6), spiked with 10 pmoles of the miSPIKE (Integrated DNA Technologies) 21-mer control RNA, M-21 nt RNA size control; (b) the example 15% denaturing PAGE electrophoresis of end linkering reaction for small RNAs from developing ovules at different DPA (0 to 6), M1–62 nt RNA size control, M2–21 nt small RNA size control; (c) 2% high-resolution agarose gel picture where RT-PCR product of and end linker ligated small RNAs of ovules was loaded, M-50 bp size ladder. Arrows indicate the small RNA fraction in (a), and linker ligated small RNA products ((b) and (c)).
