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Sigma-Aldrich

SYBR® Green Extract-N-Amp™ Tissue PCR Kit

sufficient for 100 extractions, sufficient for 100 amplifications

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About This Item

UNSPSC Code:
41106303
NACRES:
NA.55

usage

sufficient for 100 amplifications
 sufficient for 100 extractions
 sufficient for 100 reactions

Quality Level

200

feature

dNTPs included
hotstart

technique(s)

PCR: suitable

storage temp.

−20°C

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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.

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 Hot Start® SYBR® Green PCR ReadyMix™ for highly specific PCR amplification and quantitation of genomic DNA
  • Safe – no organic extraction with hazardous chemicals

Components

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.

Principle

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.

Other Notes

For additional information, please see www.sigma-aldrich.com/extract-n-amp.

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 Life Technologies

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Signal Word

Danger

Hazard Classifications

Aquatic Chronic 2 - Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - Skin Sens. 1 - STOT SE 3

Target Organs

Respiratory system

Storage Class Code

10 - Combustible liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

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Linda Doan et al.
Journal of visualized experiments : JoVE, (11)(11), doi:10-doi:10 (2008-12-11)
Genomic detection of DNA via PCR amplification and detection on an electrophoretic gel is a standard way that the genotype of a tissue sample is determined. Conventional preparation of tissues for PCR-ready DNA often take several hours to days, depending
Haroldo A Toque et al.
PloS one, 8(8), e72277-e72277 (2013-08-27)
Elevated arginase (Arg) activity is reported to be involved in diabetes-induced vascular endothelial dysfunction. It can reduce L-arginine availability to nitric oxide (NO) synthase (NOS) and NO production. Akita mice, a genetic non-obese type 1 diabetes model, recapitulate human diabetes.
Thomas Prukop et al.
Journal of neuroscience research, 98(10), 1933-1952 (2020-06-27)
Charcot-Marie-Tooth disease 1 A (CMT1A) is caused by an intrachromosomal duplication of the gene encoding for PMP22 leading to peripheral nerve dysmyelination, axonal loss, and progressive muscle weakness. No therapy is available. PXT3003 is a low-dose combination of baclofen, naltrexone
Modesto Rojas et al.
PloS one, 8(12), e84357-e84357 (2013-12-21)
Diabetic retinopathy, a major cause of blindness, is characterized by increased expression of vascular endothelial growth factor (VEGF), leukocyte attachment to the vessel walls and increased vascular permeability. Previous work has shown that reactive oxygen species (ROS) produced by the
Ilya Chumakov et al.
Orphanet journal of rare diseases, 9, 201-201 (2014-12-11)
Charcot-Marie-Tooth disease type 1A (CMT1A) is the most common inherited sensory and motor peripheral neuropathy. It is caused by PMP22 overexpression which leads to defects of peripheral myelination, loss of long axons, and progressive impairment then disability. There is no

Articles

Genomic DNA Sample Purification and Quality Assessment

The availability of simple methods for purification of DNA and RNA has greatly facilitated the analysis and characterization of the genome and gene expression. There is a demand to isolate DNA and RNA rapidly and conveniently from a variety of cellular sources, including cells and tissues from mammalian, plant and bacterial cultures.

Protocols

Rapid Genotyping of Mouse Tissue Using Extract-N-Amp Tissue PCR Kit

Genomic detection of DNA via PCR amplification and detection on an electrophoretic gel is a standard way that the genotype of a tissue sample is determined. Conventional preparation of tissues for PCR-ready DNA often take several hours to days, depending on the tissue sample. The genotype of the sample may thus be delayed for several days, which is not an option for many different types of experiments. Here we demonstrate the complete genotyping of a mouse tail sample, including tissue digestion and PCR readout, in one and a half hours using our SYBR Green Extract-N-Amp™ Tissue PCR Kit.

SYBR® Green Extract-N-Amp™ Tissue PCR Kit Protocol

The SYBR® Green Extract-N-Amp™ Tissue PCR Kit 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.

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