Animal Tissue RNA Purification Kit For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Animal Tissue RNA Purification Kit SKU 25700 Format Spin Column Size 50 Preps Quantity Trouver un distributeur Contact Overview Details Documentation Citations Aperçu du produit This kit is suitable for the isolation of total RNA from a range of animal tissues such as liver and spleen, as well as difficult fibrous tissues such as heart, muscle, intestine, etc. Briefly, the tissue of interest is first lysed using Buffer RL, followed by treatment with the provided Proteinase K which aids in the removal of the various proteins present in fiber-rich tissues including collagen, contractile proteins, and connective tissues. The purified RNA is of the highest quality and purity, with excellent RIN values and A260/A280, and is suitable for downstream applications including qRT-PCR, RT-PCR, microarrays, NGS, and more. The kit purifies all sizes of RNA from large mRNA, lncRNA down to microRNA (miRNA) in the same fraction without the requirement of phenol. Isolate all RNA sequences at an equal rate irrespective of size. Moreover, when the RNA sequences are small (e.g. miRNA), the column binds small RNAs regardless of their GC content. Détails Supporting Data Previous Figure 1. High Quality of Isolated RNA with Complete Size Range. Unlike most competitor’s kits, Norgen's Animal Tissue RNA Purification Kit allows for the isolation of all sizes of RNA, from the very large RNA down to the microRNA without the use of phenol. Total RNA was isolated from 25 mg samples of rat muscle using Norgen’s Animal Tissue RNA Purification Kit and a leading competitor's kit. Five microliters of the 50 µL RNA eluted from each kit were then resolved on a 1.2% formaldehyde-agarose gel. As it can be seen, only Norgen’s kit was able to isolate the full diversity of RNA species, including all the small RNA species Figure 2. High Quality RNA Purification from a Diversity of Tissues. Norgen’s 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 Norgen’s 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 Norgen’s 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, Norgen’s 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 Norgen’s 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, Norgen’s 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®. Norgen’s 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 Norgen’s 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, Norgen’s 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, Norgen’s kit isolated much higher yields of microRNA than the competitor’s 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. Next Click for expanded view Kit Specifications Maximum Column Binding Capacity 50 μg Maximum Column Loading Volume 650 μL Size of RNA Purified All sizes, including small RNA < 200 nt Time to Complete 10 Purifications 50 minutes Average Yield Liver (10mg) Kidney (10mg) Spleen (10mg) Heart (10mg) Muscle (10mg) 30 μg 25 μg 15 μg 10 μg 5 μg Maximum Amount of Starting Material Heart Kidney Liver Muscle Spleen 30 mg 15 mg 15 mg 30 mg 15 mg Storage Conditions and Product Stability The DNAse I should be stored at -20°C upon arrival. The Proteinase K should be stored at -20°C upon arrival and after reconstitution. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date shipment. Component Cat. 25700 (50 preps) Buffer RL 30 mL RNase-Free Water 40 mL Wash Solution A 38 mL Enzyme Incubation Buffer A 6 mL Elution Solution A 6 mL Proteinase K 2 vials DNase I 1 vial Mini Spin Columns 50 Collection Tubes 100 Elution Tubes (1.7 mL) 50 Product Insert 1 Documentation PROTOCOLS (25700) Animal Tissue RNA Purification Kit - Protocol (50 prep) SAFETY DATA SHEETS (25700) Animal Tissue RNA Purification Kit - Safety Data Sheet FLYERS Flyer RELATED BLOGS 3 Reasons why Norgen’s Industry-Leading Patented RNA Purification Technology is Best-in-Class How to Reduce False Negative SARS-CoV-2 Results with Silicon Carbide Related Products Leukocyte RNA Purification Kit microRNA Purification Kit RNase-Free DNase I Kit
Animal Tissue RNA Purification Kit For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Animal Tissue RNA Purification Kit SKU 25700 Format Spin Column Size 50 Preps Quantity Trouver un distributeur Contact