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DNA & RNA Purification

SYBR Green Extract-N-Amp Tissue Kit


Description:
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.
  • 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

The SYBR Green Extract-N-Amp Tissue PCR Kit offers an innovative extraction system that eliminates the need for either long enzymatic digestions or homogenization. The product includes a specially formulated Hot Start SYBR Green PCR ReadyMix™ for amplification and quantitation directly from the extract.

Procedure: 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. The extract is ready for real-time PCR in any plate-based real-time thermal cycling system in less than
15 minutes!

Application: Ideal for genotyping, gene copy number experiments, and amplifying and quantifying DNA from multiple tissue sample types.

Watch the video protocol Rapid Genotyping of Mouse Tissue Using Sigma's Extract-N-Amp Tissue PCR Kit


Watch the full video protocol here
Video brought to you by JoVE.com
SYBR Green Extract-N-Amp Tissue Flow Chart


Incubate with Extraction Solution for 10 minutes at room temperature.

Heat at 95 °C for 3 minutes.



Add Neutralization Solution.



Mix aliquot with Extract-N-Amp™ SYBR® Green PCR ReadyMix™ and primers.


Transfer to thermal cycler.

PCR.


Beneficial Features
Speed From tissue to qPCR in less than 15 minutes!
Convenient No long enzymatic digestions needed.
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.

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Order Information
Product Code Product Name Number of Extractions Number of Amplifications Technical Bulletin.pdf
XNATG SYBR® Green Extract-N-Amp Tissue PCR Kit 100 100 TB
XNATRG SYBR® Green Extract-N-Amp Tissue PCR Kit 1000 1000 TB

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Distinguish differences in gene copy number
SYBR Green Extract-N-Amp Distinguishes Between 1-2 Gene Copies

Figure 1. SYBR Green Extract-N-Amp Used to Distinguish between 1-2 Gene Copies. Extracts were prepared following the standard protocol for the SYBR Green Extract-N-Amp Tissue PCR Kit from 8 mouse tails that contained either a single or double copy of the Diap2 gene. Two single-gene PCR reactions were run on each extract using the 2x Extract-N-Amp SYBR Green PCR ReadyMix.

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Flexible enough for a wide variety of tissue types
SYBR Green Extract-N-Amp PCR Data for a Variety of Tissue Types

Figure 2. Quantitative PCR of DNA isolated from a variety of tissue types and sources. The Extract-N-Amp Tissue PCR Kit was used to extract and amplify genomic DNA from various sources. The extracted DNA was then amplified using the specially formulated Hot Start SYBR Green PCR ReadyMix.

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Highly sensitive SYBR Green PCR ReadyMix™
Highly Sensitive SYBR Green PCR ReadyMix

Figure 3. Quantitative PCR of DNA isolated from a series of decreasing extract dilutions. DNA was extracted from a mouse tail snip following the standard protocol for the SYBR Green Extract-N-Amp Tissue PCR Kit. The extract was diluted in decreasing three-fold increments, from 100% extract to 0.14% extract. As depicted above, message can be clearly detected even with the most dilute extract.

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