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Simple and Rapid Isolation of Plant RNA,,, Using the Eppendorf Perfect RNA, Eukaryotic, Maxi Kit

nd 600 g RNA were yielded from 1 g, 5 g, and 10 g leaves, respectively. Two g sprouts yielded 170 g RNA. When 2 g source material from a BMS maize cell line was used (suspension culture), the yield was 600 g RNA. Purity was determined by examining the A260/280 ratio. All samples had ratios between 1.9 and 2.05, demonstrating a high degree of purity. Figure 1 shows that the Perfect RNA isolation procedure produces intact and non-degraded RNA. Figure 2 represents successful RT-PCR amplification of a 2331 bp and an 891 bp-specific gene fragment of Arabidopsis.

Figure 1: Two g total RNA from Arabidopsis leaves (lane 2), flowers (lane 3), and pods (lane 4) were run on a 1% agarose formaldehyde gel and stained with ethidium bromide. Lane 1: 0.24-9.5 kb RNA Ladder from Invitrogen/Life Technologies. RNA had been isolated using the Perfect RNA Maxi Kit. The distinct 18S and 28S rRNA bands demonstrate the excellent total RNA quality. Mitochondrial and chloroplast ribosomal RNA are also visible. Figure 2: Ethidium bromide-stained 1% agarose gel. The gel demonstrates the successful amplification of the 2331 bp Acc. BAB08877 and 891 bp Acc. BAB01090 Arabidopsis gene fragment. RNA was isolated using the Perfect RNA Eukaryotic Maxi Kit. Five l each of the RT reaction had been used for agarose gel electrophoresis. Lane M: 1 kb standard (Invitrogen/LifeTechnologies); Lane 1: amplified 2331 bp fragment; Lane 2: negative control without RNA; Lane 3: amplified 891 bp fragment; Lane 4: negative c
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