Catalog# : NB-24200
Price/Unit : $295.00
Unit : 20 preps
Purify RNA, microRNA, total proteins and genomic DNA from the same sample
* Purify RNA, microRNA, proteins and DNA from a single sample
* No phenol extraction step
* Fast and easy processing
* Ideal for precious, difficult to obtain or small samples
* Flexible protocol allows for total RNA purificaiton (including microRNA) or separate enrichment of microRNAs
Bio-Synthesis's All-in-One Purification Kit provides a rapid method for the isolation and purification of total RNA including microRNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants. In addition, a microRNA Enrichment Column is provided for the optional seperate purification of small RNA (<200 nt), allowing for 4-in-1 purification (large RNA, genomic DNA, microRNA and proteins). This kit is an ideal all-in-one solution for researchers studying systems biology, including those who are interested in the interactions of multiple disciplines including RNA interference, genomics, epigenomics, transcriptomics and proteomics. Bio-Synthesis's All-in-One Purification Kit is especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as needle biopsies and single foci from cell cultures, as well as for mutant analysis, RNA interference studies and pathogen detection. Furthermore, analysis will be more reliable since the RNA, DNA and proteins are derived from the same sample without any fractionation, thereby eliminating inconsistent results. The purification procedure is very rapid, allowing for the isolation of large RNA, genomic DNA, microRNA and proteins from a single sample in less than 40 minutes. The isolated macromolecules are of the highest purity and can be used in a number of different downstream applications.
Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Briefly, the process involves first lysing the cells or tissue of interest with the provided Lysis Solution. Ethanol is then added to the lysate, and the solution is loaded onto a spin-column. Under these conditions only the RNA and DNA will bind to Bio-Synthesis's resin, while the proteins are removed in the flowthrough (BIND 1). An additional protocol utilizing the microRNA Enrichment Column is provided if the user wishes to further separate the large (>200 nt) and the small/microRNA (<200 nt). Next, the bound RNA is washed with the provided RNA Wash Solution to remove impurities, and the purified RNA is eluted with the RNA Elution Solution (WASH ELUTE 1A). The remaining bound genomic DNA is then washed with the provided gDNA Wash Buffer to remove any remaining trace levels of RNA, and the gDNA is then eluted with the gDNA Elution Buffer (WASH ELUTE 1B). The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).
The proteins that are present from the initial flowthrough can now be loaded directly onto an SDS-PAGE gel for visual analysis. Alternatively, the protein samples can be further purified using the spin columns provided with the kit. After the RNA and gDNA has been eluted from the column, the flowthrough is then pH adjusted and loaded back onto the column in order to bind the proteins that are present (BIND 2). The bound proteins are washed with the provided wash buffer (WASH 2), and are then eluted such that they can be used in downstream applications (ELUTE 2).
The purified RNA, DNA and proteins are of the highest quality and can be used in a number of downstream applications.
FEATURES AND BENEFITS* No phenol - Isolate RNA, DNA and proteins without the use of phenol or chloroform
* Complete column purification Isolate total or micro-enriched RNA- Two different protocols are provided to isolate either total RNA, including all sizes of RNA from large rRNA to microRNA, or separately isolate microRNA species (<200>200 nt)
* Reduce variability - RNA, DNA and proteins are isolated from a single sample with no splitting of the lysate, thus reducing inconsistent results and variability.
* Isolate from small samples - Sequential isolation of RNA, DNA and protein from a single sample. Ideal for precious, difficult to obtain or small samples.
* Rapid procedure - Isolate total RNA, genomic DNA and total proteins OR microRNA, large RNA, genomic DNA and proteins from a single sample in < 40 minutes.
* Isolate a diversity of RNA species - All sizes of RNA are isolated, from large mRNA down to microRNA, either separately or combined in a total RNA fraction.
* Process a wide range of sample types - Isolate total RNA, genomic DNA and proteins from cultured animal cells , tissue, blood, bacteria, yeast, fungi and plants.
APPLICATIONS
RNA
* 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
* microRNA cloning
* PCR-based virus detection
* PCR-based viable bacteria detection
Genomic DNA
* PCR and Quantitative PCR
* Sequencing
* DNA methylation studies
* Southern blotting
* Microarrays
* SNP analysis
* PCR-based virus detection/identification
* PCR-based bacteria detection/identification
Protein
* SDS-PAGE analysis
* Western blots
* Mass spectrometry
Figure 1. Sequential Isolation of microRNA, Large RNA, Genomic DNA and Proteins from 1 x 106 HeLa Cells. Panels A and B show the separate isolation of large RNA and microRNA from 2 different samples of HeLa cells. Panel A is a 1X MOPS 1% agarose gel and Panel B is a 10% urea-PAGE gel. In both gels, Lane M is Bio-Synthesis's 1Kb RNA Ladder, Lanes 1 and 2 contain 3 µL out of the 50 µL elutions of the large RNA fraction, and Lanes 3 and 4 contain 3 µL out of the 50 µL elutions of the microRNA fractions. Panel C is a 1% agarose gel showing the gDNA isolated from the same 2 HeLa cell samples. Lane M is Bio-Synthesis's UltraRanger DNA Ladder and Lanes 1 and 2 contain 10 µL of each of the 100 µL elutions. Panel D is a 12% SDS-PAGE gel that contains the proteins that were isolated from the 2 HeLa cell samples. Lane M is a protein ladder and Lanes 1 and 2 contain 10 µL of the 100 µL elution of proteins that had been column purified. The large RNA, microRNA, gDNA and proteins are all intact and of the highest integrity and quality. Furthermore, the kit allows for the successful separate isolation of microRNA (<200>200nt).
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