Figure 2. High Quality RNA Purification from a Diversity of Tissues. Norgens Animal Tissue RNA Purification Kit isolates high-quality RNA from a diversity of tissues including both tissues with high RNA content and fibrous tissues. Total RNA was successfully isolated from various tissue samples including 10 mg of spleen (Panel C), 10 mg of kidney (Panel D), and 25 mg of fiber-rich muscle (Panel B) using Norgens Animal Tissue RNA Purification Kit and a leading competitor's kit (Panel A). All RNA samples were eluted in a 50 µL volume. One microliter of each of the purified RNA samples was then analyzed using an Agilent BioAnalyzer on an RNA Nano 6000 Chip. All RNA samples resolved well in the Bioanalyzer suggesting high RNA quality. Moreover, small RNA could be observed in all the RNA samples isolated using Norgens kit (Panels B-D, circled), whereas no small RNA was present in the samples isolated using a competitors kit (Panel A).

Figure 3. Total RNA with High Yield of microRNA Species. Unlike most competitor kits that exclude RNA species of size < 200 nt, Norgens Animal Tissue RNA Purification Kit isolates the complete size range of total RNA including small RNAs. Total RNA was isolated from 25 mg of a rat muscle sample using Norgens Animal Tissue RNA Purification Kit and a leading competitor's kit as shown in Figure 1. Two microliters of the 50 µL isolated RNA were then subjected to a 20 µL reverse transcription using miR-21 stem-loop reverse primer or oligo dT. Two microliters of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the rat miR-21 (Panel A) and the beta-actin transcripts (Panel B), respectively. In both graphs, the blue lines correspond to Norgen isolated-RNA and the red lines correspond to competitor-isolated RNA. As it can be seen, Norgens kit isolated much higher yields of microRNA, as indicated by the lower Ct value of the blue line in Panel A. The kits isolated similar yields of mRNA as seen by the similar Ct values in Panel B. NTC corresponds to no-template control.

Figure 4. Compatibility with Tissues Stored in RNAlater®. Norgens Animal Tissue RNA Purification Kit can isolate total RNA from both flash frozen tissues and tissues stored in RNA preservation media such as RNAlater®. Total RNA was isolated from 10 mg of a hamster kidney sample, either flash frozen or stored in RNAlater®, using Norgens Animal Tissue RNA Purification Kit and a leading competitor's kit. Three microliters of the 50 µL isolated RNA were then subjected to a 20 µL reverse transcription using miR-21 stem-loop reverse primer or oligo dT. One microliter of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the hamster miR-21 (Panel A) and the beta-actin transcripts (Panel B), respectively. In both graphs, the blue lines correspond to Norgen isolated-RNA and the red lines correspond to competitor-isolated RNA, both from RNAlater®-preserved samples. The green lines correspond to Norgen isolated-RNA from flash frozen samples. As it can be seen, Norgens kit isolated similar amounts of both mRNA and microRNA from either frozen or RNAlater®-preserved samples, as shown by the similar Ct values of the blue and green lines. In addition, Norgens kit isolated much higher yields of microRNA than the competitors kit, as indicated by the lower Ct value of the blue and green lines in Panel A. The kits isolated similar yields of mRNA as seen by the similar Ct values in Panel B. NTC corresponds to no-template control.