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 p
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
'"/>Source:
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