Total RNA Purification Plus Kit
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For research use only and NOT intended for in vitro diagnostics.
Aperçu du produit
This kit purifies total RNA including miRNA from biological samples and employs an extra column for the efficient removal of contaminating gDNA, thereby replacing the enzymantic DNase step.
Efficiently extract total RNA from a range of samples including cells, bacteria, yeast, virus and bodily fluids including plasma/serum, blood, saliva, CSF and more. Extract high quality and purity RNA with excellent RIN values and A260/A280 suitable for downstream applications including qRT-PCR, RT-PCR, microarrays, NGS and more.
The kit purifies all sizes of RNA from large mRNA, lncRNA down to microRNA (miRNA) in the same fraction without the requirement of phenol. Isolate all RNA sequences at an equal rate irrespective of size. Moreover, when the RNA sequences are small (e.g. miRNA), the column binds small RNAs regardless of their GC content.
Détails
Supporting Data
Figure 1. Genomic DNA Removal from HeLa cells. Norgen's Total RNA Purification Plus Kit is unique in that it contains a Genomic DNA Removal Column, allowing for gDNA removal without the use of enzymes. HeLa cells were either passed through the gDNA Removal Column, or received no gDNA removal treatment. The Genomic DNA Removal Column successfully removed gDNA from both 1 million HeLa cells (Panel A) and 1x105 HeLa cells (Panel B). For both Panels A and B, the left image displays the RNA run on a 1.2% EtBR-agarose gel, to better visualize any gDNA contamination. The right image displays a 1.0% formaldehyde-agarose gel to allow for better denaturation of RNA in the sample. For higher inputs, as in image A, the Genomic DNA Removal Column also resulted in higher RNA yields than the non-treated samples.
Figure 2. Delta Ct Values Observed Between Different gDNA Removal Techniques. Genomic DNA is well known to contaminate RNA samples, and subsequently RT-qPCR results. Therefore, to determine the amount of genomic DNA contamination in RNA samples, an RT-qPCR can be used to compare Ct values observed with and without reverse transcription of the RNA sample. This is known as the Delta Ct. One million and 1x105 HeLa cells were either passed through the gDNA Removal Column or subjected to DNase-treatment using Norgen's RNase-Free DNase I (Cat# 25710). The gDNA Removal column eliminated gDNA similar to a DNase treatment. For higher inputs (i.e. 1 Million HeLa cells), the gDNA Removal Column successfully removed more gDNA contamination from the RNA samples than the DNase-treatment. When inputs were lowered, the amount of genomic DNA contamination in the DNase-treated sample was similar to the sample passed through the gDNA Removal Column. Both methods work well for removing gDNA contamination in RNA samples.
Kit Specifications
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Maximum Binding Capacity Per Column
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Up to 50 μg RNA
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Maximum Loading Volume Per Spin Column
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650 μL
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Size of RNA Purified
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All sizes, including < 200 nt
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Time to Complete 10 Purifications |
25 minutes
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Maximum Amount of Starting Material
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Liver Tissue
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20 mg
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Heart Tissue
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5 mg
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Kidney Tissue
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20 mg
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Brain Tissue
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25 mg
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Lung Tissue |
20 mg
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Spleen Tissue
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20 mg
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Whole Blood
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100 µL
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Hela Cells
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3 x 106
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CHO |
3 x 106
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Yeast |
1 x 108 cells
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E.coli |
1 x 109 CFU/mL
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Bacteria | 1 x 109 cells | ||
Plasma/Serum | 200 µL | ||
Fungi |
50 mg
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Plant Tissue |
50 mg
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Viruses |
≤100 µL of viral suspension
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Average Yield
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All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
Component | Cat. 48300 (50 preps) | Cat. 48400 (100 preps) |
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Buffer RL | 40 mL | 2 x 40 mL |
Wash Solution A | 38 mL | 2 x 38 mL |
Elution Solution A | 6 mL | 2 x 6 mL |
RNA Purification Columns | 50 | 100 |
Genomic DNA Removal Columns | 50 | 100 |
Collection Tubes | 100 | 200 |
Elution Tubes (1.7 mL) | 50 | 100 |
Product Insert | 1 | 1 |