GenElute™ Mammalian Total RNA Miniprep Kit
SIGMA/RTN70 - sufficient for 70 purifications
Synonym: GenElute™ Mammalian RNA Kit; animal total RNA purification kit; cell RNA extraction kit; cell RNA isolation kit; cell total RNA purification kit; tissue total RNA purification kit; total RNA extraction kit; total RNA isolation kit; total RNA purification kit; Gen Elute
Product Type: Chemical
High Quality RNA from various tissues and cells.
Formaldehyde-agarose gel and Northern blot of total RNA purified with GenElute™ Mammalian Total RNA Purification Kit. 2 μg of each RNA analyzed on a 1.2% agarose gel containing 0.6 M formaldehyde.
Formaldehyde-agarose gel and Northern blot of total RNA purified with GenElute™ Mammalian Total RNA Purification Kit. 2 μg of each RNA analyzed on a 1.2% agarose gel containing 0.6 M formaldehyde.
RT-PCR detection with RNA from ≥100 cells.
HeLa cells were diluted to give 0, 100, 300, or 1,000 cells per tube, and RNA was prepared with the GenElute™ Mammalian Total RNA Isolation Kit. Duplicate 10 μL samples (20%) of each preparation were treated with Amplification Grade DNase I (Cat. No. AMPD1). One of each pair was reverse transcribed with eAMV™ Reverse Transcriptase (Cat. No. A4464; + lanes). The other was incubated under the same conditions, but without the reverse transcriptase (– lanes). PCR was performed with 2 μL (10%) from each reaction, G3PDH primers, and Taq DNA Polymerase (Cat. No. D1806). One-fifth of each PCR product was fractionated on a 1.5% agarose gel, and photographed after staining for 30 minutes with SYBR® Green I (Cat. No. S9430). RT-PCR products are clearly visible for all reactions with reverse transcriptase added (+ lanes), except those with no cells (0). The control lanes contain 1 ng of total RNA prepared form the same cell line. No PCR products are visible for reactions without reverse transcriptase added (– lanes), demonstrating that RT-PCR products with reverse transcriptase are from RNA and not from residual, contaminating DNA.
HeLa cells were diluted to give 0, 100, 300, or 1,000 cells per tube, and RNA was prepared with the GenElute™ Mammalian Total RNA Isolation Kit. Duplicate 10 μL samples (20%) of each preparation were treated with Amplification Grade DNase I (Cat. No. AMPD1). One of each pair was reverse transcribed with eAMV™ Reverse Transcriptase (Cat. No. A4464; + lanes). The other was incubated under the same conditions, but without the reverse transcriptase (– lanes). PCR was performed with 2 μL (10%) from each reaction, G3PDH primers, and Taq DNA Polymerase (Cat. No. D1806). One-fifth of each PCR product was fractionated on a 1.5% agarose gel, and photographed after staining for 30 minutes with SYBR® Green I (Cat. No. S9430). RT-PCR products are clearly visible for all reactions with reverse transcriptase added (+ lanes), except those with no cells (0). The control lanes contain 1 ng of total RNA prepared form the same cell line. No PCR products are visible for reactions without reverse transcriptase added (– lanes), demonstrating that RT-PCR products with reverse transcriptase are from RNA and not from residual, contaminating DNA.
Total RNA was prepared from 100, 200, 300, 400, 500, and 600 μL of HeLa cell lysate (~ 1, 2, 3, 4, 5, & 6 x 106 cells) with either Sigma’s GenElute Mammalian Total RNA Isolation Kit or with a RNA kit from Supplier Q. The amount of RNA recovered in two 50 μL elutions was determined by absorbance at 260 nm. Note that the capacity of Supplier Q′s column is ~100 μg, as claimed in their technical bulletin, whereas that of the Sigma column is ~150-160 μg. Quality of RNA prepared with the 2 kits is similar (data not shown).
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
| greener alternative category | Aligned  , |
| greener alternative product characteristics | Designing Safer Chemicals Learn more about the Principles of Green Chemistry . |
| Quality Level | 200 |
| storage temp. | 15-25°C |
| sustainability | Greener Alternative Product |
| technique(s) | RNA purification: suitable |
| usage | sufficient for 70 purifications |
| Application: | GenElute™ Mammalian Total RNA Miniprep Kit has been used to isolate total RNA: • from snap-frozen preoptic area of rat brain • from central part of the mantle tissue containing gonads of Mytilus trossulus mussel • from murine liver |
| Application: | The purified RNA is ready for reverse transcription and PCR, labeling and microarray analysis, and other common applications. Note that RNA shorter than 200 nucleotides in length, such as tRNA, 5S rRNA, and 5.8S rRNA, is not recovered efficiently under the conditions used with this kit. |
| Biochem/physiol Actions: | Samples are lysed and homogenized in guanidine thiocyanate and 2-mercaptoethanol to release RNA and inactivate RNases. Lysates are spun through a filtration column to remove cellular debris and shear DNA. The filtrate is then applied to a high capacity silica column to bind total RNA, followed by washing and elution. Up to 150 μg of total RNA can be recovered per prep in 100 μl of water. The purified RNA is ready for Northern blots (Fig. 1), RT-PCR (Fig. 2) and other common applications. |
| Biochem/physiol Actions: | The GenElute Mammalian Total RNA Miniprep Kit combines silica membrane technology with a convenient spin column format for a rapid bind, wash, and elute method to prepare high quality total RNA. |
| Features and Benefits: | • Purifies total RNA from up to 107 cells or 40 mg of tissue per prep • Yields up to 150 μg of pure, concentrated total RNA per prep • Recover RNA from as few as 100 cells • Simple and efficient–12 to 18 preps in 30 minutes • Faster than gravity flow anion exchange methods • No cumbersome steps associated with resins and magnetic slurries • 40% more purifications per kit than the leading supplier |
| Features and Benefits: | • Yields up to 150 μg of pure, concentrated total RNA per prep • Purifies total RNA from up to 107 cells or 40 mg of tissue per prep • Recover RNA from as few as 100 cells |
| General description: | Click here to view the kit protocol |
| General description: | Sigma′s GenElute Mammalian Total RNA Miniprep Kit provides a simple and convenient way to isolate total RNA from mammalian cells and tissues. Protocols are provided for cells, tissues, and fibrous tissues. These protocols differ in their cell lysis and disruption conditions. Once the RNA is bound to the GenElute Binding Column, the purification procedure is the same for all starting materials. For fibrous tissues the kit must be used with proteinase K (Cat. No. P4850) to ensure effective cell disruption. The kit combines the advantages of a silica-based system with a microspin format and eliminates the need for cesium chloride gradients, alcohol precipitation, and hazardous organic compounds such as phenol and chloroform. Cells or tissues are lysed and homogenized in a buffer containing guanidine thiocyanate to ensure thorough denaturation of macromolecules and inactivation of RNases. Addition of ethanol causes RNA to bind when the lysate is spun through a silica membrane in a microcentrifuge tube. After washing to remove contaminants, RNA is eluted in 50-100 μL of Elution Solution. Up to 150 μg of total RNA can be isolated in less than 30 minutes. If all traces of DNA contamination must be eliminated, further treatment with DNase I is recommended. DNase I digestion can be performed while the RNA is bound to the GenElute Binding Column using the On-Column DNase I Digestion Set (DNASE10 and DNASE70). Alternatively, for more stringent removal of contaminating DNA, the final RNA preparation can be treated with Amplification Grade DNase I (AMPD1). |
| General description: | We are committed to bringing you Greener Alternative Products, which adhere to one or more of The 12 Principles of Greener Chemistry. This product has Inherently Safer Chemistry, compared to the standard use of phenol and chloroform to perform DNA extractions. |
| Legal Information: | GenElute is a trademark of Sigma-Aldrich Co. LLC |
| Other Notes: | For additional information, please see www.sigma-aldrich.com/tot. |
| Other Notes: | The GenElute™ Mammalian Total RNA Purification Kit combines silica-membrane technology with a convenient spin column format for a rapid bind, wash, and elute method to prepare high quality total RNA. |
| Packaging: | Components included:• Qty in kit:4 Part number: T5449 Component description: Collection Tubes, 2 mL • Qty in kit:1 Part number: CP9346 Component description: GenElute Filtration Columns • Qty in kit:1 Part number: W3136 Component description: Wash Solution 1 • Qty in kit:1 Part number: E8024 Component description: Elution solution • Qty in kit:1 Part number: CP9471 Component description: GenElute Miniprep Binding Columns • Qty in kit:1 Part number: M3148 Component description: 2-Mercaptoethanol • Qty in kit:1 Part number: L8285 Component description: Lysis solution • Qty in kit:1 Part number: W3261 Component description: Wash solution 2 Concentrate |
| Symbol |     GHS05,GHS06,GHS08,GHS09 |
| Signal word | Danger |
| Hazard statements | H301 + H331 - H310 - H314 - H317 - H361d - H373 - H410 |
| Precautionary statements | P202 - P273 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 |
| Hazard Codes | T,N |
| Risk Statements | 23/24/25-32-34-43-48/22-50/53 |
| Safety Statements | 26-36/37/39-45-61 |
| RIDADR | UN 3316 9 |
| WGK Germany | WGK 3 |
| Supplemental Hazard Statements | EUH032 |
| Storage Temp. | 15-25°C |
| UNSPSC | 41105501 |