Sample Preparation |
Purification of Total RNA
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Poster from RNA Society (714k)
Applications
- Purification of RNA from yeast and filamentous fungi suitable for cDNA synthesis
and microarray gene expression analysis (Figs. 1 and 2).
The MasterPure™ Yeast RNA Purification Kit provides all of the reagents
needed to purify RNA from yeast cell types (including Candida, Saccharomyces, Schizosaccharomyces)
and filamentous fungi. The kit uses a rapid salting-out process1 to
remove contaminating macromolecules, avoiding toxic organic solvents, bead-beating,
and spheroplasting.2
Benefits
- Avoids hot acid phenol.
- Faster than spheroplasting.
- Higher quality RNA than bead-beating.
- No extra enzymes or equipment to purchase.
- Yields 25-50 µg RNA from 1 ml of mid-log S.cerevisiae.
References
- Miller, S.A. et al., (1988) Nucleic Acids Res. 16, 1215.
- Hoffman, L.M. and Jarvis, B.W. (2004) EPICENTRE Forum 11(2),
4.
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| Figure 1. Microarray analysis of yeast RNA purified with
the MasterPure™ Yeast RNA Purification Kit. The RNA was reverse
transcribed, labeled, and hybridized to a S. cerevisiae DNA microarray.
The cDNA from cells under oxidative stress was labeled with Cy5, and cDNA
from control cells was labeled with Cy3. |
Figure 2. Purity of RNA obtained using the MasterPure™ Yeast
RNA Purification Kit. The RNA used in Fig. 1. was stored at -20° C
for two months before analysis on an Agilent Technologies 2100 Bioanalyzer®.
The electrophoretogram demonstrates the high-purity, intact RNA. |
MasterAmp™ High Fidelity RT-PCR Kit
MasterAmp™ RT-PCR Kit for High Sensitivity
MonsterScript™ 1st-Strand cDNA Synthesis Kit
MonsterScript™ Reverse Transcriptase
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