REDExtract-N-Amp™ Blood PCR Kit
SIGMA/XNABS - sufficient for 10 extractions, sufficient for 10 amplifications
Product Type: Chemical
Direct sequence from PCR products generated using the Extract-N-Amp™ Blood Kit.
A 547 bp product for human surfactant protein B was generated using the Extract-N-Amp Blood PCR kit. The product was sequenced directly using BigDye™ terminator chemistry. Sequencing reactions were resolved on an ABI 3100.
Note: Some PCR products require further clean-up prior to sequencing. The GenElute™ PCR Clean-Up Kit (Cat. No. NA1020) is recommended.
A 547 bp product for human surfactant protein B was generated using the Extract-N-Amp Blood PCR kit. The product was sequenced directly using BigDye™ terminator chemistry. Sequencing reactions were resolved on an ABI 3100.
Note: Some PCR products require further clean-up prior to sequencing. The GenElute™ PCR Clean-Up Kit (Cat. No. NA1020) is recommended.
Direct sequence from PCR products generated using the Extract-N-Amp™ Blood Kit.
A 547 bp product for human surfactant protein B was generated using the Extract-N-Amp Blood PCR kit. The product was sequenced directly using BigDye® terminator chemistry. Sequencing reactions were resolved on an ABI 3100.
Note: Some PCR products require further clean-up prior to sequencing. The GenElute™ PCR Clean-Up Kit (Cat. No. NA1020) is recommended.
A 547 bp product for human surfactant protein B was generated using the Extract-N-Amp Blood PCR kit. The product was sequenced directly using BigDye® terminator chemistry. Sequencing reactions were resolved on an ABI 3100.
Note: Some PCR products require further clean-up prior to sequencing. The GenElute™ PCR Clean-Up Kit (Cat. No. NA1020) is recommended.
PCR analysis of genomic DNA isolated from blood using Sigma′s Extract-N-Amp™ Blood PCR Kit.
Extract-N-Amp™ Blood PCR Kit used to isolate genomic DNA from fresh, 7 day old, and frozen blood. DNA was extracted and neutralized from 10 μL of whole blood in 5 minutes at room temperature using the REDExtract-N-Amp™ Blood PCR kit. The PCR products were then amplified using the specially formulated Hot Start PCR mix included in the kit. PCR products generated are 1.8 kb for carnitine palmitoyltransferase II, 1.3 kb for a mitochondrial DNA control region, 547 bp for human surfactant protein B, and 320 bp for the 5′ untranslated region of human major histocompatibility complex class II.
Extract-N-Amp™ Blood PCR Kit used to isolate genomic DNA from fresh, 7 day old, and frozen blood. DNA was extracted and neutralized from 10 μL of whole blood in 5 minutes at room temperature using the REDExtract-N-Amp™ Blood PCR kit. The PCR products were then amplified using the specially formulated Hot Start PCR mix included in the kit. PCR products generated are 1.8 kb for carnitine palmitoyltransferase II, 1.3 kb for a mitochondrial DNA control region, 547 bp for human surfactant protein B, and 320 bp for the 5′ untranslated region of human major histocompatibility complex class II.
Stability of Extract-N-Amp™ Blood Extracts.
Blood was drawn from 2 human volunteers into vacutainer tubes containing EDTA. Extractions were performed in duplicate providing 4 samples total. Half the extracts were stored at 4 °C (recommended storage conditions) and the other half at 37 °C (accelerated storage). Samples were removed at various time intervals for testing. Stability was determined by monitoring yield from quantitative PCR using an ABI 7700 instrument. The DNA standards used for the quantitative PCR were generated from the same blood draw as the test samples, purified using the GenElute Blood Genomic DNA Kit (Cat. No. NA2000) and stored as single aliquots at -20 °C. The PCR products were generated using primers for a 547 bp product from human surfactant protein B (SPB) [Lin, Z. and Floros, J., BioTechniques, 29(3), 460 (2000)]. The results clearly show no loss of amplification of the SPB PCR product even after storage at 37 °C for 6 months. Similar results were obtained with storage at 4 °C.
Blood was drawn from 2 human volunteers into vacutainer tubes containing EDTA. Extractions were performed in duplicate providing 4 samples total. Half the extracts were stored at 4 °C (recommended storage conditions) and the other half at 37 °C (accelerated storage). Samples were removed at various time intervals for testing. Stability was determined by monitoring yield from quantitative PCR using an ABI 7700 instrument. The DNA standards used for the quantitative PCR were generated from the same blood draw as the test samples, purified using the GenElute Blood Genomic DNA Kit (Cat. No. NA2000) and stored as single aliquots at -20 °C. The PCR products were generated using primers for a 547 bp product from human surfactant protein B (SPB) [Lin, Z. and Floros, J., BioTechniques, 29(3), 460 (2000)]. The results clearly show no loss of amplification of the SPB PCR product even after storage at 37 °C for 6 months. Similar results were obtained with storage at 4 °C.
Representative image of product packaging.
Representative image of product packaging.
| color | red |
| feature | dNTPs included |
| hotstart | |
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | −20°C |
| technique(s) | PCR: suitable |
| usage | sufficient for 10 amplifications |
| sufficient for 10 extractions | |
| sufficient for 10 reactions |
| Application: | REDExtract-N-Amp™ Blood PCR Kit has been used to extract genomic DNA from blood samples, peripheral blood leukocytes, parasites and endocervical swab eluates of mice. It has also been used to perform polymerase chain reaction (PCR) using oligonucleotide primers. |
| Features and Benefits: | • Efficient 8 minute prep allows greater speed and throughput • No need for organic extraction, centrifugation or alcohol precipitation • Hot start antibody for highly specific PCR amplification of genomic DNA • Can be used with whole blood or blood cards • Extract stable at 4 °C for at least 6 months • Compatible with any format (single tube, 96-well, etc.) • Simple, 3 step procedure with no special equipment required • REDExtract-N-Amp™ requires no loading buffers or tracking dyes required for gel analysis • PCR products are suitable for direct cloning and sequencing |
| General description: | The REDExtract-N-Amp™ Blood PCR Kits contain all of the reagents necessary to rapidly extract host genomic DNA from whole blood and amplify targets of interest by direct PCR (polymerase chain reaction). This novel extraction system eliminates the need for any type of purification, organic extraction, centrifugation, heating, filtration or alcohol precipitation. The kit also includes a PCR ReadyMix™, specially formulated for amplification directly from the extract. This formulation uses an antibody based hot start, for specific amplification. The REDExtract-N-Amp Blood PCR Kit contains a tracking dye that allows for convenient direct loading of PCR reactions onto agarose gels for analysis. |
| Legal Information: | Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser′s own internal research. No other patent rights (such as 5′ Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. Antibody licensed for in vitro research use under U.S. Patent No. 5,338,671 and 5,587,287, and corresponding patents in other countries. |
| Legal Information: | ReadyMix is a trademark of Sigma-Aldrich Co. LLC |
| Legal Information: | REDExtract-N-Amp is a trademark of Sigma-Aldrich Co. LLC |
| Legal Information: | Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,789,224, 5,618,711, 6,127,155 and claims outside the US corresponding to expired US Patent No. 5,079,352. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. |
| Other Notes: | For additional information, please see www.sigma-aldrich.com/ext. |
| Principle: | Genomic DNA is extracted from 10 μl of whole blood by simply adding the Extraction Solution and incubating for 5 minutes at room temperature. The Neutralization Solution is added to the extract to counteract inhibitory substances prior to PCR. A portion of the DNA extract is then added to the specially formulated PCR ReadyMix. |
| Symbol |  GHS05 |
| Signal word | Danger |
| Hazard statements | H314 |
| Precautionary statements | P280 - P305 + P351 + P338 - P310 |
| Hazard Codes | C |
| Risk Statements | 34 |
| Safety Statements | 26-36/37/39-45 |
| RIDADR | UN 3316 9 |
| Storage Temp. | −20°C |
| UNSPSC | 12352200 |
| Components | Lysis Solution for Blood; Neutralization Solution for Blood; REDExtract-N-Amp™ PCR ReadyMix™ for Blood |