Figure 2
Analysis of products formed in RT reactions with or without the use of modified TS oligos. (A) Comparison of background amplification between small biological samples of different sizes and negative control using standard TS oligo. cDNA synthesis for different quantities of starting material after TS-PCR (lanes 1, 4 and 7). Sfi I digestion of negative control (lane 9) and samples representing 500 and 100 tomato glands (lanes 3, 6). Lanes 2, 5, and 8 are mock restriction digestions without enzyme. (B) Comparison of cDNA synthesis performed using TS oligo vs. iso 3 TS oligo. Negative control cDNA synthesis reactions without RNA but with standard TS oligo (lane 1) or with iso3TS oligo (lane 3). cDNA synthesis with tomato gland RNA sample using TS oligo (lane 2) and iso3TS oligo (lane 4). (C) Comparison of Sfi I digested tomato glandular trichome cDNA after TS-PCR cDNA synthesis using standard TS oligo vs. iso 3 TS oligo. Tomato gland cDNA synthesized using TS oligo (lane 1) and after Sfi I digestion (lanes 2 and 3), or using iso3TS oligo (lane 4) and digested with Sfi I (lanes 5 and 6). (D) Size distribution and concentration of cDNA samples produced using the iso 3 TS oligo with larger RNA samples. Lane 1: cDNA from rhizome tips of scouring rush (Equisetum hymale); lane 2: cDNA from rhizome elongation zones of scouring rush; lane 3: cDNA from rhizome tips of red rice (Oryza longistaminata); lane 4: cDNA from rhizome elongation zones of red rice; L: 500 ng of 1 kb DNA ladder (NEB).