Figure 1.  Consistent Yield of High Quality RNA with Complete Size Range without the Use of Phenol.   Norgen's Total RNA Purification 96-Well Kit allows for the consistent isolation of high quality RNA, with complete size range from the very large RNA down to small RNA without the use of phenol. Total RNA was isolated from 0.5 mL aliquotes of an overnight culture of E. coli (~5 x 10⁸ cells/mL) using Norgen’s Total RNA Purification 96-Well Kit in 16 replicates. Five microliters of the 75 µL isolated RNA was resolved on a 1.2% formaldehyde agarose gel. All 16 replicates showed both high yield and high quality. In addition, all replicates showed effective recovery of all sizes of RNA including the small RNA (arrow).

Figure 2. High Quality of RNA from a Diverse Range of Inputs.  Norgen’s Total RNA Purification 96-Well Kit allows for RNA isolation from a wide range of species and tissue types. Total RNA was isolated from 8 mg of brain (Lanes A), kidney (Lanes B), liver (Lanes C), lung (Lanes D) and spleen tissue (Lanes E), 7.5 x 10⁵ HeLa (Lanes F) and CHO cells (Lanes G), and 5 x 10⁸ bacteria (Lanes H) using Norgen’s Total RNA Purification 96-Well Kit. Five microliters of the 75 µL isolated RNA was resolved on a 1.2% formaldehyde agarose gel. From observing the formaldehyde-agarose gel, it can been seen that Norgen’s kit can be used to successfully isolate total RNA, including small RNA species, from a broad range of sample types.

Figure 3. Isolation of Both Large and Small RNA.  Norgen’s Total RNA Purification 96-Well Kit allows for consistent isolation of both large and small RNA. Total RNA was isolated from samples of 5 x 10⁵ HeLa cells using Norgen’s Total RNA Purification 96-Well Kit. Aliquots of each total RNA sample were then used in 2 different RT-qPCR reaction. Ten microliters of the 75 µL isolated RNA was subjected to a 20 µL reverse transcription using miR-21 stem-loop reverse primer or oligo dT. Three microliters of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the human miR-21 (Panel A) and the S15 transcripts (Panel B), respectively. Both the mRNA and microRNA were amplified in a consistent manner from all the samples, with low variability of the Ct values. Thus both types of RNA are being consistently isolated using this kit.

Figure 4. Great Isolation Sensitivity.   Norgen’s Total RNA Purification Kit allows sensitive RNA extraction from even less than 10 cells. RT-qPCR was used to detect mRNA isolated from various input amounts of HeLa cells, from 100,000 down to a single cell, using Norgen’s Total RNA Purification 96-Well Kit. Ten microliters of the 75 µL isolated RNA was then subjected to a 20 µL reverse transcription using oligo dT primer. Three microliter of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the human S15 transcripts. The resulting Ct values were plotted against the input cell number. Total RNA was isolated and detected linearly from as little as a single HeLa cell.