Catalog# : NB-24300
Price/Unit : $620.00
Unit : 2 plates
For rapid high-throughput purification of total RNA including microRNA
* True total RNA isolation including miRNA and siRNA
* No phenol extraction step
* Highly sensitive
* Fast and easy processing
Bio-Synthesis's Total RNA Purification 96-Well Kit provides a rapid method for the high-throughput isolation and purification of total RNA using a 96-well format in 30 minutes. 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. Total RNA can be isolated from a broad range of sample sources including cultured cells, tissues, blood, serum, plasma, bacteria, yeast, fungi, plants and viruses. The kit is highly sensitive, as RNA can be isolated from as little as a single cell.
Purification is based on 96-well spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Purification can be performed on either a vacuum manifold or using centrifugation. Briefly, the cells or tissue of interest is first lysed using the provided Lysis Solution, ethanol is added and the RNA is bound to Bio-Synthesis's 96-well filter plates (BIND). Under these conditions only the RNA will bind to Bio-Synthesis's resin while most of the contaminating cellular proteins are removed in the flowthrough or retained on top of the resin. The bound RNA is then washed to remove any remaining impurities (WASH). Lastly, the purified total RNA is eluted into 75 µL of the provided Elution Buffer or water (ELUTE).Please see procedure flowchart to the right.
Bio-Synthesis's proprietary resin provides superior affinity to the full size range of RNA molecules, resulting in large and small RNA (miRNA) purification with excellent consistency and sensitivity. 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, expression array assays, miRNA cloning and Next Generation Sequencing.
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)
* Isolate RNA from a wide range of samples - RNA can be isolated from cultured animal cells, small tissue samples, blood, plasma, serum, bacteria, yeast, fungi, plants and viruses
* No phenol:chloroform extractions - Total RNA, including all small RNA species, is isolated without the use of harmful chemicals such as phenol or chloroform
* Isolate RNA from very small samples - Total RNA has been isolated and detected from as little as a single animal cell
* Fast and easy processing - 96-well plates can be rapidly processed in 30 minutes using either a vacuum manifold or centrifugation format
* Recovered RNA is suitable for many 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 assays, miRNA cloning and Next Generation Sequencing
APPLICATIONS
Purified RNA is of the highest quality and can be used in a number of downstream applications including:
* 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
* miRNA from plasma for discovery
* microRNA cloning
* PCR-based virus detection
* PCR-based viable bacteria detection
Figure 1. Consistent Yield of High Quality RNA with Complete Size Range without the Use of Phenol. Bio-Synthesis'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 Bio-Synthesis'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. Bio-Synthesis'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 Bio-Synthesis'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 Bio-Synthesis'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. Bio-Synthesis'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 Bio-Synthesis'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.
Total RNA Purification 96-Well Kit Components1. Lysis Solution
2. Wash Solution
3. Elution Buffer
4. 96-Well Filter Plate
5. Adhesive Tape
6. Elution Plate
7. Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. 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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