Description
General description
The SYBR® Green Extract-N-Amp™ Tissue PCR kit is a direct PCR (polymerase chain reaction) kit that contains all the reagents needed for rapid extraction, amplification and detection of genomic DNA from mouse tails and other animal tissues, buccal swabs, hair shafts, and saliva.
Application
Extract-N-Amp Video Protocol
SYBR® Green Extract-N-Amp™ tissue PCR kit has been used for DNA extraction from embryo head of male Pmp22 transgenic Sprague–Dawley rats and in genotyping.
This direct PCR (polymerase chain reaction) kit is also suitable for gene copy number experiments and amplifying and quantifying DNA from multiple tissue sample types.
Biochem/physiol Actions
DNA is rapidly extracted from a tissue by incubating the sample with a mixture of the Extraction Solution and the Tissue Preparation Solution at room temperature for 10 minutes. After a 3-minute heat denaturing step, an equal volume of Neutralization Solution B is added to the extract to neutralize inhibitory substances and the extract is ready for real-time PCR in any plate-based real-time thermal cycling system. An aliquot of the neutralized extract is then combined with the Extract-N-Amp™ SYBR® Green PCR ReadyMix™ and user-provided PCR primers.
Features and Benefits
Novel – all liquid, single-step extraction of genomic DNA for quantitative PCR (qPCR)
Fast – tissue to qPCR in 15 minutes
Convenient – no long enzymatic digestions and no column purifications
Simple – rapid, easy-to-follow protocol
Sensitive – specially formulated HOTSTART® SYBR® Green PCR ReadyMix™ for highly specific PCR amplification and quantitation of genomic DNA
Safe – no organic extraction with hazardous chemicals
Other Notes
For additional information, please see www.sigma-aldrich.com/extract-n-amp.
Kit contains Extraction Solution, Neutralization Solution, Tissue Preparation Solution, and Extract-N-Amp SYBR Green PCR ReadyMix™. The Extract-N-Amp SYBR Green PCR ReadyMix is a 2X reaction mix containing SYBR Green, buffer, salts, dNTPs, Taq polymerase and JumpStart™ Taq antibody. It is optimized specifically for use with the extraction reagents and contains JumpStart Taq antibody for hot start PCR to enhance specificity and SYBR Green I to act as a nonspecific reporter for real-time PCR.
Legal Information
Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,994,056 and 6,171,785. 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 (such as apparatus or system claims in US Patent No. 6,814,934) 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.
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.
Extract-N-Amp is a trademark of Sigma-Aldrich Co. LLC
HOTSTART is a registered trademark of Molecular BioProducts, Inc.
JumpStart is a trademark of Sigma-Aldrich Co. LLC
ReadyMix is a trademark of Sigma-Aldrich Co. LLC
SYBR is a registered trademark of Thermo Fisher Scientific or its subsidiaries
