Catalog# : NB-25700
Price/Unit : $320.00
Unit : 50 preps
For purification of total RNA (including microRNA) from all types of tissues including fiber-rich tissues
* No need for phenol
* Isolate total RNA, including microRNA, from animal tissues
* Purified RNA is of the highest quality and integrity
Bio-Synthesis's Animal Tissue RNA Purification Kit provides a rapid method for the isolation and purification of total RNA from all types of animal tissue samples, including fiber-rich tissues such as muscle and heart. The kit can isolate RNA from fresh or flash-frozen tissues, as well as tissues stored in RNA preservation reagents such as RNAlater®. Bio-Synthesis's Animal Tissue RNA Purification Kit is provided with Proteinase K, which aids in the removal of the various proteins present in fiber-rich tissues including collagen, contractile proteins and connective tissues. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), without the use of inhibitory phenol or chloroform.
Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Briefly, the tissue of interest is first lysed using the Lysis Solution, followed by treatment with the provided Proteinase K. Ethanol is then added, and the RNA is bound to Bio-Synthesis's column (BIND). Under these conditions only the RNA will bind to Bio-Synthesis's resin while most of the contaminating cellular proteins and contaminants are removed in the flowthrough or retained on top of the resin. At this point an optional on-column DNase digestion can be performed using the provided DNase to remove any traces of DNA which may co-purify with the RNA. The bound RNA is then washed to remove any remaining impurities (WASH). Lastly, the purified total RNA is eluted into 50 µL of the provided Elution Buffer or water (ELUTE).Please see procedure flowchart to the right.
The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
FEATURES AND BENEFITS
* Isolate a diversity of RNA species - All RNA species can be isolated, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA)
* Fast and easy processing - Rapid spin-column format allows for the processing of 10 samples in 50 minutes
* No phenol:chloroform extractions - Total RNA is isolated without the use of harmful chemicals such as phenol or chloroform
* Isolate total RNA from diverse range of tissue - Total RNA has been isolated and detected from regular high RNA content tissues such as liver and spleen and difficult to extract fibrous tissues such as heart and muscle
* Compatible with RNA Preservation Reagents - Total RNA has been isolated and detected from tissues stored in RNAlater®
* Recovered RNA is suitable for downstream applications - Purified RNA can be used in a number of downstream applications including real-time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array analysis requiring the use of intact RNA
APPLICATIONS
* Quantitative, real-time RT-PCR for both large mRNA and small RNA including miRNA
* RT-PCR for both large mRNA and small RNA including miRNA
* Northern blotting
* RNase protection
* Primer extension
* Expression array assays
* Next Generation Sequencing for RNA and miRNA
* microRNA cloning
Figure 1. High Quality of Isolated RNA with Complete Size Range. Unlike most competitor’s kits, Bio-Synthesis's Total 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 Bio-Synthesis's Animal Tissue RNA Purification Kit and a leading competitors kit. Five microliters of the 50 µL RNA eluted from each kit was then resolved on a 1.2% formaldehyde-agarose gel. As it can be seen, only Bio-Synthesis'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. Bio-Synthesis'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 Bio-Synthesis's Animal Tissue RNA Purification Kit and a leading competitors 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 Bio-Synthesis's kit (Panels B-D, circled), whereas no small RNA was present in the samples isolated using a competitors kit (Panel A).
Figure 3. Tissue Total RNA with High Yield of microRNA Species. Unlike most competitor kits that exclude RNA species of size <200>
Figure 4. Compatibility with Tissues Stored in RNAlater®. Bio-Synthesis'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 Bio-Synthesis's Animal Tissue RNA Purification Kit and a leading competitors kit. Three microliters of the 50 µL isolated RNA was 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 Bio-Synthesis's isolated-RNA and the red lines correspond to competitor-isolated RNA, both from RNAlater®-preserved samples. The green lines correspond to Bio-Synthesis's isolated-RNA from flash frozen samples. As it can be seen, Bio-Synthesis'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, Bio-Synthesis'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 non-template control.
Components
1. Lysis Solution
2. RNase-Free Water
3. Wash Solution
4. Enzyme Incubation Buffer
5. Elution Solution
6. Proteinase K
7. DNase I
8 Mini Spin Columns
9 Collection Tubes
10 Elution tubes
11 Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. The DNAse I should be stored at -20°C upon arrival. The Proteinase K should be stored in aliquots at -20°C upon reconstitution. These reagents should remain stable for at least 1 year in their unopened containers.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.
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