Extract-N-Amp™ Tissue PCR Kit

Description

The Extract-N-Amp™ Tissue PCR Kits provide all the reagents necessary to rapidly extract DNA from a wide variety of cells and tissues and amplify targets of interest by PCR (Figure 1).

  • Novel – single-step extraction of genomic DNA to PCR
  • Fast – tissue to PCR in 15 minutes
  • Convenient – no long enzymatic digestions

The Extract-N-Amp Tissue PCR Kit offers a novel extraction system that eliminates the need for either long enzymatic digestions or homogenization. The product includes a specially formulated hot start PCR ReadyMix™ for amplification directly from the extract. Extract-N-Amp products are Sigma Advanced Technology certified.

Watch the full video protocol here
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Procedure

Genomic DNA is extracted from a tissue sample that has been incubated in Tissue Preparation Solution and Extraction Solution for 10 minutes at room termperature. The sample is heated to 95 °C for 3 minutes and then mixed with a third solution to neutralize inhibitory substances prior to PCR. An aliquot of the DNA extract is then added directly to the optimized PCR mix supplied.

Application

Perfect for genotyping.
Watch the video protocol: Rapid Genotyping of Mouse Tissue Using Sigma's Extract-N-Amp Tissue PCR Kit

Beneficial Features

Starting Material Mouse tail (0.5 to 1.0 cm)
tissue (2 to 10 mg)
buccal (10 µL)
Speed Extraction of genomic DNA for PCR in 15 minutes
Convenient No long enzymatic digestions
Safe No phenol/chloroform or other hazardous materials
Specific Hot Start antibody for highly specific PCR amplification of genomic DNA

Pick the formulation that is right for you

The Extract-N-Amp Tissue PCR Kits include a specially formulated hot start PCR ReadyMix for amplification directly from the extract.

The PCR ReadyMix comes in two formulations:

  • REDExtract-N-Amp PCR Ready Mix – contains an inert dye that acts as a tracking dye and allows for convenient loading of PCR reactions onto agarose gels for analysis.
  • Extract-N-Amp PCR Ready Mix – no dye contained.

Order Information

Extract-N-Amp Tissue PCR Kits

Product No. Description Number of Extractions Number of Amplifications Technical Manual (PDF)
XNAT2 Extract-N-Amp Tissue PCR Kit 100 100 PDF
XNAT2R Extract-N-Amp Tissue PCR Kit 1000 1000 PDF


REDExtract-N-Amp Tissue PCR Kits

Product No. Description Number of Extractions Number of Amplifications Technical Manual (PDF)
XNAT-10RXN
REDExtract-N-Amp Tissue PCR Kit (contains REDTaq) 10 10 PDF
XNAT-100RXN REDExtract-N-Amp Tissue PCR Kit (contains REDTaq) 100 100 PDF
XNAT-1000RXN REDExtract-N-Amp Tissue PCR Kit (contains REDTaq) 1000 1000 PDF

Kit Contents

Extract-N-Amp Kit REDExtract-N-Amp Kit
  • Extraction Solution
  • Tissue Preparation Solution
  • Neutralization Solution B
  • Extract-N-Amp PCR Reaction Mix
  • Extraction Solution
  • Tissue Preparation Solution
  • Neutralization Solution B
  • REDExtract-N-Amp PCR Reaction Mix

Sample Data

PCR analysis of genomic DNA extracted from various samples using Sigma's Extract-N-Amp Tissue PCR Kit.

Figure 1. 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 PCR ReadyMix. The products generated are 1181 bp for the Interleukin 1 b gene in mouse and 1820 bp for the Carnitine palmitoyltransferase II in human. Markers are PCR Marker (Product No. P9577).

 

Sequence determination for 1181 bp Interleukin-1b mouse-tail PCR product.

Figure 2. The PCR product was purified with the GenElute™ PCR Clean-Up kit. The DNA Extraction and PCR were performed using Sigma's Extract-N-Amp Tissue PCR Kit. The sequence was obtained using ABI Big Dye™ Terminator Chemistry and the same primers as for the original PCR. Reaction products were resolved on an ABI 310.

 

Stability of DNA in mouse-tail extracts.

Figure 3. Mouse-tail samples were extracted according to the procedure in the Technical Bulletin for the Extract-N-Amp Tissue PCR Kit. Extracts were stored at 37 °C instead of the recommended 4 °C to test accelerated storage and stability. Samples were removed at various time intervals for testing. Stability was determined by monitoring yield for quantitative PCR using SYBR® Green on an ABI Prism® 7700 instrument. Robust signals were obtained even after storage for 10 weeks.
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