C·Prep Phenol/Trizol is a supplemental kit which is used in conjunction with acid guanidinium thiocyanate-phenol-chloroform lysis buffers, such as Trizol. Phenol/chloroform extraction is robust and works with nearly any sample type.
This product improves on the standard phenol/chloroform extraction method. RNA yield is 30% better on average because no chloroform or layer separation is needed, the process time is reduced to less then 15 minutes, the method becomes easy to automate, trace phenol contamination is eliminated which significantly 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||General purpose RNA isolation. Works with any sample type and provides the highest purity and yield. The main disadvantage is that acid guanidinium-phenol lysis buffers are dangerous. Many sequencing cores will not recommend methods that use Trizol and similar lysis buffers because trace phenol contamination interferes with sequencing. The carbon technology solves this problem, trace DNA and phenol contamination are both eliminated in RNA samples.|
|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||All samples. Plant samples have separate protocol.|
|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.|
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