Description
General description
The REDExtract-N-Amp™ Seed PCR Kits provide all the reagents necessary to rapidly extract DNA from a wide variety of plant seeds and amplify targets of interest by direct PCR (polymerase chain reaction). A novel extraction method eliminates the need for long enzymatic digestions. The kit also includes a specially formulated hot start PCR reaction mix for amplification directly from the extract. The REDExtract-N-Amp PCR ReadyMix contains an inert dye that acts as a tracking dye and allows for convenient loading of PCR reactions onto agarose gels for analysis.
Application
REDExtract-N-Amp™ seed PCR Kit has been used for DNA extraction and to amplify hygromycin resistance gene.(hpt) by polymerase chain reaction (PCR).
Biochem/physiol Actions
The kit comes with validated protocols to extract and amplify genomic DNA from corn, wheat, sunflower, cotton, soybean, sorghum, canola, and Arabidopsis seeds. Additional protocols are available. In a typical procedure, genomic DNA is extracted from a sample that has been ground and incubated in the seed preparation solution and extraction solution for 10 minutes at room temperature. The sample is heated to 95 °C for 3 minutes and then mixed with a third solution to neutralize inhibitory substances prior to PCR. A portion of the DNA extract is then added to a PCR reaction containing primers and the Extract-N-Amp PCR Reaction Mix (included in the kit) and PCR is initiated.
Features and Benefits
Perfect for quick genotyping results
Rapid extraction of genomic DNA for PCR in 15 minutes
No long enzymatic digestions
Hot Start antibody for highly specific PCR amplification of genomic DNA
Compatible with a wide variety of seed sources
Extracts are stable at 4 °C for at least 6 months, allowing for multiple reassays
Phenol/chloroform extraction or alcohol precipitation not required
Column purification or centrifugation not required
Other Notes
For additional information, please see www.sigma-aldrich.com/extract-n-amp.
Legal Information
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. SYBR Green nucleic acid stain is licensed from Molecular Probes, Inc. under US Patent No. 5,436,134 for use in Real-Time PCR by end-users for research and development only. SYBR is a registered trademark of Molecular Probes, Inc.
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.
Extract-N-Amp is a trademark of Sigma-Aldrich Co. LLC
REDExtract-N-Amp is a trademark of Sigma-Aldrich Co. LLC
