CarbonPrep for animal tissues is a ready-to-use kit which comes with buffers formulated for use with animal tissues such as kidney, liver, lung, spleen, pancreas, ect. The protocol can be scaled to different input sizes but the standard input size is 8 mg of animal tissue.
Purpose | RNA isolation/purification from tissues such as liver, kidney, ect. |
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Format and Technology | Magnetic Beads; Carbon Technology |
Processing Time | 15 min |
Type of RNA Purified | Total RNA (includes ribosomal RNA, mRNA, and small RNAs) |
Application | Northern, dot, and slot blotting, end-point RT-PCR, quantitative, real-time RT-PCR, next-generation sequencing |
Processing | Manual or automated. Manual protocols use a water bath to release RNA, while automated protocols use robotic agitation to release RNA. |
Main Sample Type | Tissue samples such as liver, kidney, and lung. For difficult samples such as bacterial biofilms we recommend the Phenol/Trizol kit. All C·Prep kits can be reconfigured into other C·Prep kits with changes to the buffers see the kit contents document under protocols for details. |
Elution Volume | 30-100 µl |
Sample Size | 8 mg of tissue is the standard input and reagent consumption scales with the amount of tissue. Less tissue may be used and the reagent use scales accordingly meaning that if less tissue is used the kit will be good for more than the listed number of reactions and vice versa. |
Kit Storage | Room Temperature, one vial should be stored at 4° C |
Genomic DNA contamination | Carbon technology is >100x more selective than silica for RNA. In routine extractions gDNA contamination is typically ~0.5% – 1% of the sample making DNase cleanup unnecessary for routine PCR applications. If your application requires even higher purity, optimization of the protocol for your specific sample can result in <0.1% gDNA contamination. |
A comparison of extraction technologies using mouse kidney tissue is presented below. Technologies were compared by a third party at Wayne State University using the method presented in the following paper: “Padhi, B. K., Singh, M., Huang, N., & Pelletier, G. (2016). A PCR-based approach to assess genomic DNA contamination in RNA: Application to rat RNA samples. Analytical Biochemistry, 494, 49–51.” https://doi.org/10.1016/j.ab.2015.10.012
As compared to silica-based products, carbon has virtually no DNA contamination and does not require DNase cleanup. This is a huge advantage since DNase cleanup increases time and cost and damages RNA. During the COVID-19 pandemic, labs with at-home saliva tests cited lower DNA contamination in samples as an important factor for lower COVID-19 detection limits.
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Safety data sheets for all materials included in kit: