Catalog# : NB-23600
Price/Unit : $240.00
Unit : 50 preps
For rapid and efficient clean-up and concentration of total RNA, including microRNA, without phenol
* Clean-up and concentrate total RNA in minutes
* No phenol or chloroform extractions
* Clean-up RNA from contaminants including enzymes, RNases and nucleotides
* Rapid spin-column protocol
* Purified RNA is fully compatible with all downstream applications
Bio-Synthesis's RNA Clean-Up and Concentration Kit provides a rapid method for the purification, cleanup and concentration of up to 35 μg of RNA isolated using different methods including phenol/guanidine-based protocols and from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. The kit can concentrate as much as 35 µg of RNA to as little as picograms of RNA. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other reaction components such as proteins, RNases and nucleotides without the use of phenol or chloroform.
Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Briefly, the RNA samples or reactions containing RNA are mixed with Binding Solution, ethanol is 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 proteins and nucleotides are removed in the flowthrough. The bound RNA is then washed to remove any remaining impurities (WASH). Lastly, the purified total RNA is eluted into 20 - 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. Bio-Synthesis's RNA Clean-Up and Concentration Kit purifies RNA with minimal amounts of DNA contamination. An optional protocol is provided for maximum removal of residual DNA that may affect sensitive downstream applications such quantitative PCR.
FEATURES AND BENEFITS
* Efficient clean-up from various enzymatic reactions - RNA clean-up from upstream enzymatic applications such as labeling, DNase treatment and in vitro transcription. Remove proteins, RNases, DNases, nucleotides, etc.
* Clean-up and concentration of RNA isolated by different methods - RNA that has been isolated using various methods, including phenol-chloroform extractions or alcohol precipitations, can be processed with this kit
* Fast and easy processing - Rapid spin-column format allows for the processing of 10 samples in 20 minutes
* No phenol:chloroform extractions - Bio-Synthesis's RNA Clean-Up and Concentration Kit purifies RNA without the use of harmful chemicals such as phenol or chloroform.
* Isolate all small RNA molecules - All small RNA species can be isolated including microRNA, siRNA, tRNA, and 5S rRNA
* Recovered RNA is suitable for downstream applications - Purified RNA can be used in a number of downstream applications including end-point or quantitative reverse transcription PCR, Northern blotting, and expression array analysis
APPLICATIONS
Purified RNA is of the highest quality and can be used in a number of downstream applciations including:
* Bioanalyzer
* 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. Effective Clean-Up to Produce High Quality Total RNA with Complete Size Diversity. Bio-Synthesis's RNA Clean-Up and Concentration Kit effectively cleans up RNA isolated from phenol-based extractions without the loss of RNA diversity by retaining all RNA species including small RNAs. Total RNA was isolated from 5 x 108 E. coli using a competitor’s phenol-based RNA extraction reagent. The resulting RNA was then purified using Bio-Synthesis's RNA Clean-Up and Concentration Kit. As controls, total RNA was extracted using both Bio-Synthesis's Total RNA Purfication Kit (#17200, no phenol required) and the phenol-based RNA extraction reagent only without any clean-up. Resolution of 7 µL of the 50 µL purified RNA on a 1X MOPS, 1.5% formaldehyde-agarose gel showed the RNA extracted with phenol was successfully cleaned-up by Bio-Synthesis's RNA Clean-Up and Concentration Kit without the loss of small RNA species.
Figure 2. Effective Clean-Up to Produce High Quality Total RNA Compatible to Bioanalyzer Analysis. Total RNA was isolated from 0.75 million HeLa cells using a competitor’s phenol-based RNA extraction reagent. The resulting RNA was then purified using Bio-Synthesis's RNA Clean-Up and Concentration Kit. As controls, total RNA was extracted using both Bio-Synthesis's Total RNA Purfication Kit (#17200, no phenol required) or the phenol-based RNA extraction reagent only without clean-up. Resolution of 1 µL of the 50 µL purified RNA on an Agilent RNA Nano 6000 Chip showed that without clean-up, the RNA sample isolated using the phenol-based RNA extraction reagent only resolved poorly on a bioanalyzer. In contrast, RNA extracted with the phenol-based RNA extraction reagent followed by clean-up by Bio-Synthesis's RNA Clean-Up and Concentration Kit resolved properly on the bioanalyzer showing excellent quality.
RNA Clean-Up and Concentration Kit Contents1. Spin Column
2. Binding Solution
3. Wash Solution
4. Elution Buffer
5. Micro Spin Columns
6. Collection Tubes
7. Elution Tubes
8. 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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