C·Prep Phenol/Trizol is a supplemental kit which is used in conjunction with acid guanidinium thiocyanate-phenol-chloroform lysis buffers, such as Trizol, to improve upon this difficult but powerful RNA purification method.
Compared to the standard phenol/chloroform extraction, RNA yield is 30% better on average, the process time is reduced to less then 15 minutes, the method becomes easy to automate, phenol contamination is reduced which improves RNA sequencing results, and the protocol can be stopped half-way to create carbon-RNA complexes which stabilize RNA at room temperature for shipping and storage for >30 days.
|Purpose||RNA isolation/purification where acid-guanidinium-phenol lysis buffers are used. This includes Trizol, Qiazol, TriReagent and other similar reagents, we recommend using Trizol from Millipore-Sigma. The input samples are typically bacterial biofilms, bone samples, and other difficult tissues where an aggressive lysis buffer is needed.|
|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||Any sample as long as acid-guanidinium-phenol lysis buffers are used. 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||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.|