Navigation Links
Improve RNA Isolation for Array Analysis and More

RiboPure Technology - For Tissue, Cells, Blood, Bacteria and Yeast


Researchers performing array analysis have found that a combination of phenol based and solid phase RNA extraction methods often provide superior array data than either method alone (1-3). Ambion's new line of RiboPure RNA Isolation Kits are the first kits that combine these RNA extraction methods. Four different kits are now available for the isolation of high quality total RNA from a variety of samples:

RiboPure Kit for isolating RNA from animal and plant tissues and cells

RiboPure-Yeast Kit for isolating RNA from yeast cells

RiboPure-Bacteria Kit for isolating RNA from microbes

RiboPure-WBC Kit for RNA isolation from blood samples


High Quality RNA Isolation
The RiboPure RNA Isolation Kits combine a single reagent homogenization and organic extraction step with traditional glass fiber filter purification to yield exceptionally pure RNA, free of residual proteins and lipids. The RiboPure procedure is especially suited for processing problematic samples that would otherwise clog filter cartridges (e.g. muscle, heart), samples that are difficult to disrupt (e.g. muscle, fibrous plant tissue, yeast, bacteria), or samples that are high in ribonuclease or other contaminants (e.g. spleen, pancreas, plant and neural tissues). RiboPure is the method of choice for preparing RNA for use in microarray analysis or other procedures that require RNA of exceptional purity. Each of the RiboPure Kits described below are based on this dual method technology and also include Ambion's DNA-free genomic DNA removal system, making them ideal for preparation of RNA for RT-PCR based applications.


Fast Results and High Yields with Small Samples
The RiboPure Kit employs a rapid, 15 minute RNA purification procedure that combines the best features of one step organic extraction protocols and solid phase extraction techniques. From 1 to 100 mg of tissue or 100 to 107 cells can be processed per purification. Briefly, the protocol consists of disrupting samples in Ambion's RNAwiz, which contains phenol and RNase inhibitors, then adding chloroform and mixing. The sample is then centrifuged to separate organic and aqueous phases. The aqueous phase, which contains the RNA, is recovered and mixed with ethanol. This preparation is then passed over an RNA-binding silica filter. The filter is washed to remove contaminants, and the RNA is eluted in a small volume. A subsequent DNA removal step adds ~30 minutes of additional time (mostly incubation time) to the procedure. Total RNA yield is typically 80-400 g per 100 mg tissue, depending on the type of tissue processed. The RiboPure Kit contains reagents for up to 50 RNA isolations.


New! Isolate High Quality Total RNA from Yeast
In yeast, gene expression patterns can be affected during pretreatments used to disrupt the yeast cells prior to RNA isolation. The RiboPure-Yeast RNA Isolation Kit uses no enzymatic pretreatments and thus does not alter expression profiles. RiboPure-Yeast combines a simple and efficient glass bead/one step phenol mediated disruption using Ambion's Vortex Adapter followed by a glass filter based RNA purification step. After DNase treatment with the included DNA-free Reagents, the resulting RNA is exceptionally pure (figure 1) containing little to no genomic DNA (<0.05%). The kit has been used at Ambion to isolate high quality RNA from Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia pastoris, and Candida albicans. The kit provides the highest yield of any method tested from each of these species (Figure 2). The RiboPure-Yeast RNA Isolation Kit contains sufficient reagents for 50 RNA isolations each from 107 up to 3 x 108 yeast cells.

Figure 1. Yeast Total RNA Isolated Using the RiboPure-Yeast Kit. P. pastoris, S. cerevisiae RNA, C. albicans, and S. pombe RNA, isolated using the RiboPure-Yeast Kit and treated with DNase, was run on an Agilent bioanalyzer chip. The ratio of 28S:18S rRNA is given for each sample.

Figure 2. RNA Yields From 3 Yeast Species Using Ambion's RiboPure-Yeast or Competitors' Kits. RNA was isolated from the specified number of yeast cells using the manufacturer's protocol. Following purification, all RNA samples were treated with Ambion's DNA-free reagents to remove contaminating DNA.


Isolate High Quality Total RNA from Bacteria
As with yeast, bacterial gene expression patterns can also be affected during enzymatic or chemical pretreatments used to disrupt cell walls prior to RNA isolation. The RiboPure-Bacteria RNA Isolation Kit uses no enzymatic pretreatments, allowing for faster processing, which preserves RNA expression profiles. The kit utilizes the same type of protocol combining an efficient glass bead/one step phenol mediated disruption with the Vortex Adapter followed by a glass filter based RNA purification step. The kit has been used at Ambion to isolate high quality RNA from Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Campylobacter fetus, and Rhodobacter sphaeroides and is compatible with most bacterial RNA species (Figure 3). The RiboPure-Bacteria RNA Isolation Kit contains sufficient reagents for 50 RNA isolations, each from 107 up to 109 bacteria.

Figure 3. High Quality Bacterial RNA Obtained with RiboPure-Bacteria Kit. Total RNA (2 g) was separated on a 1% agarose gel in formaldehyde/MOPS buffer. Samples were heated at 80C for 10 min in denaturing agarose gel loading buffer with 100 g/l EtBr before electrophoresis at 5 V/cm.


RiboPure-WBC: Scalable Protocol for Isolating High Quality RNA from Blood
Isolating intact RNA from blood is challenging because: 1) only white blood cells (WBCs) contain appreciable amounts of RNA and WBCs make up only about 0.2% of blood cells and 2) whole blood also contains large amounts of proteins, heme, and genomic DNA, as well as intracellular and extracellular RNases. Thus, WBCs typically need to be isolated away from the rest of the blood constituents prior to RNA isolation.

The RiboPure-WBC Kit protocol consists of three steps: 1) fractionation of whole blood by centrifugation, recovery of the WBCs and RNA stabilization in Ambion's RNAlater (provided), 2) lysis of WBCs in guanidinium and initial RNA purification by phenol/chloroform extraction, and 3) final RNA purification on a glass fiber filter.

The kit provides sufficient reagents for the purification and stabilization of WBCs from 40 whole blood samples of up to 10 ml each (2.5-10 ml of whole blood can be used per isolation). The RNA purified with the RiboPure-WBC Kit can be used in the most demanding downstream applications, such as real-time RT-PCR or array analysis. Average yields of RNA range between 4-20 g from 5 ml of whole blood; the variation is due to differences in WBC numbers between samples.


back to top


Ordering Information
Cat# Product Name Size 1924 RiboPure Kit 50 purifications 1925 RiboPure-Bacteria Kit 50 purifications 1926 RiboPure-Yeast Kit 50 purifications 10024 Vortex Adapter 1 each
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. The Best Enzyme for the Toughest PCR Challenges Improved with New Hot-Start Feature
2. Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase
3. Improved Sensitivity for Staining RNA with SYBR Gold Stain
4. Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal Fibroblasts
5. Improved Vectors for PathDetect Trans-Reporting Systems
6. Improve Amplification Specificity with Hot Start PCR Enzyme
7. Improved Quantitative Selectivity of Clenbuterol in Human Urine using High Resolution on the TSQ Quantum Mass Spectrometer
8. Gene Expression Arrays: Highly Sensitive Detection of Expression Patterns with Improved Tools for Target Amplification
9. Protect RNA and Improve Target Cell Identification
10. The Worlds Best DNase Improved TURBO DNA-free
11. Improve Recovery and Reproducibility with Pre-Treatment of Analytical TSK-GEL G3000SWXL Size Exclusion Columns
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Improve RNA Isolation for Array Analysis and More

(Date:7/25/2014)... 25, 2014  Galmed Pharmaceuticals Ltd. (Nasdaq: ... the development and commercialization of a once-daily, oral therapy ... today announced financial results for the period ended June ... , Reported cash and cash equivalents totaling $37.4 ... December 31, 2013.  , Reported a net loss ...
(Date:7/25/2014)... , July 25, 2014 Sinovac Biotech Ltd. (Nasdaq: ... in China , today announced that it ... ended June 30, 2014, after market close on Thursday, August ... prior to the market opening on Friday, August 15, 2014, ... p.m. China Standard Time) to review the Company,s financial results ...
(Date:7/24/2014)... July 24, 2014 Gain recognition ... Supply Chain of the biotech industry. Nominations are ... Awards, brought to you by the Bio ... of supply chain management professionals for the past ... stakeholders of the Biotech industry – Manufacturers, Service ...
(Date:7/24/2014)... -- ECC West Africa, LLC (ECC) signed a contract in ... Hospital (UCH) Ibadan Board of Management for the first ... and operate and maintain a new, state-of-the-art Cancer Institute ... first of the 4-phase project, ECC will conduct feasibility ... Ibadan intends to finance and develop the Cancer Institute ...
Breaking Biology Technology:Galmed Pharmaceuticals Reports First Half 2014 Financial Results 2Galmed Pharmaceuticals Reports First Half 2014 Financial Results 3Galmed Pharmaceuticals Reports First Half 2014 Financial Results 4Galmed Pharmaceuticals Reports First Half 2014 Financial Results 5Galmed Pharmaceuticals Reports First Half 2014 Financial Results 6Sinovac to Host Conference Call to Report Second Quarter 2014 Unaudited Financial Results 2Call for Submission: BSMA’s Supply Chain Management Innovation Awards 2Call for Submission: BSMA’s Supply Chain Management Innovation Awards 3ECC Signs a Contract for the Finance, Design, and Construction of a Premier Cancer Institute in Ibadan, Nigeria 2
... LAFAYETTE, Ind. - Researchers are creating a system that ... reducing a car,s fuel consumption. The effort is ... National Science Foundation and the U.S. Department of Energy. ... which is developing a prototype using thermoelectric generators, or ...
... 2010 Crescent Healthcare today announced a planned expansion ... a lease to open an infusion pharmacy in Totowa, ... services to current patients in Massachusetts, Maryland, New York, ... Rhode Island. In addition to supporting ...
... Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid ... ... if (typeof(lpUnit)==,undefined,) var lpUnit=,PRN,; --> ... var shortURL = ""; BitlyCB.alertResponse = function(data) { var s = ,; var first_result; // Results ...
Cached Biology Technology:Technology uses auto exhaust heat to create electricity, boost mileage 2Technology uses auto exhaust heat to create electricity, boost mileage 3Crescent Healthcare Expands East Coast Presence 2Oncothyreon Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid Tumors 2Oncothyreon Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid Tumors 3Oncothyreon Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid Tumors 4Oncothyreon Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid Tumors 5Oncothyreon Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid Tumors 6Oncothyreon Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid Tumors 7Oncothyreon Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid Tumors 8Oncothyreon Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid Tumors 9Oncothyreon Reports Positive Data from Phase 1 Clinical Trial of PI3-Kinase Inhibitor PX-866 in Patients with Relapsed or Refractory Solid Tumors 10
(Date:7/24/2014)... YORK, NY (July 24, 2014) Scientists at ... Institute are one step closer to creating a ... a patient,s own cells. , For the ... (iPS) cells lines from skin samples of patients ... developed an accelerated protocol to induce these stem ...
(Date:7/24/2014)... -- Keryx Biopharmaceuticals, Inc. (Nasdaq:KERX) (the "Company") announced the ... Phase 3 study of Zerenex (ferric citrate), the Company,s ... of hyperphosphatemia in patients with end-stage renal disease (ESRD) ... delivery with FErric CiTrate in EsrD) was published online ... of Nephrology ( JASN ). , ...
(Date:7/24/2014)... 24, 2014) Scientists at The New York ... step closer to creating a viable cell replacement ... cells. , For the first time, NYSCF ... from skin samples of patients with primary progressive ... protocol to induce these stem cells into becoming ...
Breaking Biology News(10 mins):NYSCF scientists one step closer to cell therapy for multiple sclerosis patients 2Zerenex™ (ferric citrate) long-term Phase 3 study results published in JASN 2Zerenex™ (ferric citrate) long-term Phase 3 study results published in JASN 3Zerenex™ (ferric citrate) long-term Phase 3 study results published in JASN 4NYSCF scientists 1 step closer to cell therapy for multiple sclerosis patients 2
... , Washington, D.C. (October 5, 2010) -- The explosion ... such as cell and smart phones, Bluetooth hands-free units, ... from the development of a wide variety of integrated ... on which the communication is based. Continuing ...
... nanotubes -- long, hollow cylinders of carbon billionths of ... in nanotechnology, optics, electronics, and many other fields. The ... scientists as yet have little control over that structure, ... growth -- of the nanotubes. In fact, says chemical ...
... A research team from the University of British Columbia and ... Hospital has identified the role of a type of T ... treatment options for young patients. Also known as juvenile ... children and young adults. In patients with type 1 diabetes, ...
Cached Biology News:The effects of hydrogen on growing carbon nanotubes 2
EZ-Tn5™ Transposase is designed for use with our pMOD™ series of vectors as well as any EZ-Tn5™ transposon system....
... PBL's bioassays for interferons are cytopathic effect ... assays. Sample types include; tissue culture supernatents; ... In these assays, one unit is defined as ... reduce the cytopathic effect of viral infection by ...
HyPro 100, includes 5 x HyPro20 and HyPro Cabinet. 100 slide capacity. Active vibration system, slide rack and humidity chamber. Ideal for microarray and in situ hybridization....
... cytopathic effect inhibition assays, also known as CPE ... culture supernatents; serum from PK studies and delivery ... defined as the quantity of interferon required per ... infection by 50%. All standards used ...
Biology Products: