Column-Pure RNA Miniprep Kit
| Cat. No. | D518 | ||||||||||||
| Name | Column-Pure RNA Miniprep Kit | ||||||||||||
| Unit | 50 preparations | ||||||||||||
| Description |
abm’s Column-Pure RNA Miniprep Kit is a fast and efficient method for the isolation and purification of total RNA from mammalian cells, tissues, yeast and bacteria. The silica spin column technology allows for rapid recovery of high quality RNA that is ready for downstream applications such as RT-PCR, qPCR, cDNA libraries and Northern blotting. Key Features:
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| Note |
Additional Materials Required:
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| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. D518 |
| What type of RNA can I isolate with this kit? | |
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The Column-Pure RNA Miniprep Kit can isolate total RNA including: mRNA, rRNA, tRNA, circRNA. Very small RNAs <50nt may be lost due to the limitations of the silica spin column.
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| Is gDNA removal necessary and how does the kit remove it? | |
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gDNA removal is necessary if performing downstream applications such as RT-qPCR or RNA-seq as it can interfere with process and/or analysis. The Column-Pure RNA Miniprep Kit includes an on-column DNaseI digestion step which can effectively remove gDNA from the RNA sample.
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| How should I store RNA after extraction? | |
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RNA should ideally be stored at -80°C (long term), or 4°C short term (1-2 days). Avoid repeated freeze/thaw cycles as this can lead to degradation.
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| How can I check the RNA purity and integrity after extraction? | |
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Common RNA quality assessment methods include a nano spectrophotometer, Bioanalyzer, and denaturing agarose gel electrophoresis. A 260/280 ratio ~2.0 indicates pure RNA; Bioanalyzer RIN scores of 7-10 reflect high integrity; and a 2:1 28S:18S rRNA band ratio on gel (for eukaryotes) indicates intact RNA. |
| Does the RNA Spin Column bind DNA as well? | |
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Yes, standard spin columns bind all nucleic acids, this is why the DNaseI treatment step is important for the isolation of pure RNA.
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| Why is my RNA yield low? | |
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Possible reasons include: Low RNA content in sample, incomplete sample lysis or homogenization, sample overload leading to clogged columns, presence of inhibitors in the lysate (e.g. polysaccharides co-precipitate/elute with RNA).
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| How can I prevent contamination and degradation of my RNA sample? | |
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1. Decontaminate work surfaces and equipment such as pipettes using a 10% bleach solution, followed by 70% ethanol.
2. Use autoclaved or certified RNase-free consumables such as tubes, tips and reagents. 3. Wear clean gloves as nucleases are present on skin. 4. Keep extracted RNA samples on ice whenever possible or store at -80°C. 5. Ensure 2-mercaptoethanol is added to the Lysis Buffer as it can denature RNases. |
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