CarbonPrep for cell cultures is a ready-to-use kit which comes with buffers formulated for use with cultured cells.
This kit is shipped with buffers pre-optimized for cell cultures, which are usually washed with phosphate buffered saline (PBS) and pelleted. The buffers and protocol included with this product have been optimized for use with samples that have a high PBS and salt content.
|Purpose||RNA isolation/purification from cultured cells|
|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||Cultured cells|
|Elution Volume||30-100 µl|
|Sample Size||2-5 million cells are the standard input and reagent consumption scales with the number of cells. Fewer cells may be used and the reagent use scales accordingly meaning that if fewer cells are used the kit will be good for more than the listed number of reactions and vice versa.|
|Kit Storage||Room Temperature|
|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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