Automation

Extract-N-Amp Tissue Kit


The Extract-N-Amp™ Tissue PCR kit is a high-throughput system for the rapid extraction and subsequent amplification of genomic DNA from mouse-tail clippings (Figures 1 and 2) and other animal tissues (Figure 3) in a 96-well format. Scientists at Sigma have developed a novel extraction treatment that releases genomic DNA from tissues in minutes. Without further cleanup the DNA is ready for direct use in PCR applications. To further simplify the procedure the Extract-N-Amp Tissue PCR kit includes a specially formulated PCR ReadyMix™, which virtually eliminates any PCR optimization.

Extract-N-Amp Tissue PCR Kit, containing Sigma Advanced Technology, offers an ideal platform for automated genomic screening of tissue samples. Features of the procedure include the following:

  • The entire process from undigested mouse-tail clippings through PCR reaction setup is fully automated.
  • Rapid method – 96 tissue samples can be processed in 45 minutes.
  • No homogenization.
  • No overnight enzymatic digestion during DNA extraction.

Automation Resources

Automated methods have been developed and validated for the Extract-N-Amp Tissue PCR Kit. The following resources are available for download:


Features & Benefits

Walk-Away Automation A liquid handler, such as the Sciclone ALH 3000 Workstation, automates all aspects of the Extract-N-Amp Tissue Kit including extraction of genomic DNA and PCR reaction setup.
Rapid Procedure 96 samples of mouse tails or animal tissue can be extracted and setup for PCR in 45 minutes. There is no need for overnight enzymatic digestion or homogenization.
Enhanced Productivity Walk-away automation of genomic DNA extraction and PCR reaction setup allows you to concentrate on other aspects of your work.
Conveniently packaged with PCR Reagents Included in the kit is the Extract-N-Amp PCR ReadyMix that is a 2x reaction mixture of buffer, salts, dNTPs, JumpStart™ Taq antibody, and Taq polymerase, and is specially formulated to compensate for PCR inhibitors that may be present after extraction.
High Specificity PCR ReadyMix includes an antibody-mediated Hot Start mechanism for highly specific amplification.
Stable Extracts Once extracts are neutralized, they are stable for at least 6 months if stored at 4° C.



Extract-N-Amp Tissue
Product No. Product Name Number of Extractions Number of Amplifications
XNAT2R Extract-N-Amp Tissue PCR 1000 1000
 
REDExtract-N-Amp™ Tissue
Product No. Product Name Number of Extractions Number of Amplifications
XNATR REDExtract-N-Amp Tissue PCR 1000 1000

Kit Contents

  • Extraction Solution
  • Tissue Preparation Solution
  • Neutralization Solution B
  • REDExtract-N-Amp or Extract-N-Amp PCR Ready Mix

 



PCR analysis of genomic DNA isolated from 88 different mouse tail clippings using an automated method for the Extract-N-Amp Tissue PCR Kit.

Figure 1. Agarose gel analysis of 96 PCR samples.

DNA was extracted from 88 different mouse tail clippings (0.3-0.4 cm) using the automated Extract-N-Amp Tissue PCR procedure on the Sciclone ALH 3000. Amplification of an 1181 bp fragment of the IL-1β gene followed using 4 µl of extracted template or 4 µl of human genomic DNA controls (A12: 3.2 ng/µl, C12: 1.6 ng/µl, E12: 0.8 ng/µl, and G12: 0.2 ng/µl) in a 20 µl PCR reaction incorporating the 2X PCR Ready Mix. Lanes B12, D12, F12, and H12 represent no template controls. 6 µl of each reaction was analyzed on a 1% agarose gel.



Cross Contamination Analysis of the Automated Extract-N-Amp Tissue Method
Figure 2. Agarose gel analysis of cross-contamination test.

Mouse tail clippings (0.3-0.4 cm long) were placed in alternating wells of a 96 well plate. The plate was processed using the automated Extract-N-Amp Tissue PCR procedure on the Sciclone ALH 3000. All samples were subjected to amplification and 6 µl of the resultant products were loaded onto a 1% agarose gel. No PCR products were detected in the wells containing no tissue samples.



PCR analysis of genomic DNA isolated from various mouse tissues using an automated method for the Extract-N-Amp Tissue PCR Kit.

Figure 3. Agarose gel analysis of PCR samples from different mouse tissue extracts.

DNA was extracted from mouse liver, mouse kidney, mouse pancreas, and mouse tails using the automated Extract-N-Amp Tissue PCR procedure on the Sciclone ALH 3000. Amplification of an 1181 bp fragment of the IL-1β gene followed using 4 µl of extracted template or 4 µl of human genomic DNA controls (Lanes A12 and C12) in a 20 µl PCR reaction incorporating the 2X PCR Ready Mix. 6 µl of each reaction was analyzed on a 1% agarose gel, with mouse liver samples in lanes A1-A11, mouse kidney samples in lanes B1-B11, mouse pancreas samples in lanes C1-C11, and mouse tails in lanes D1-D11. Lanes B12 and D12 represent no template controls.

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