Automation

Extract-N-Amp Blood Kit

 

High-throughput genetic analysis of blood requires a rapid, automated solution for genomic DNA extraction. Standard methods for purifying DNA from blood can be laborious, not amenable to automation, and unnecessary for robust downstream techniques such as PCR. The Extract-N-Amp Blood PCR Kit is a high throughput system for the rapid extraction and subsequent amplification of genomic DNA from whole blood in a 96-well automated format (Figures 1 and 2). Just place the blood samples on the deck of the liquid handler and start the instrument. A platform such as the Sciclone ALH 3000 Workstation from Caliper Life Sciences can complete the extraction and PCR reaction setup of 96 samples in just 25 minutes.

Extract-N-Amp Blood is a novel extraction system that eliminates the need for any type of purification, organic extraction, centrifugation, heating, filtration, or alcohol precipitation. A specially formulated PCR ReadyMix™ compensates for any inhibitors of PCR that may remain after the extraction, which allows for amplification directly from the extract. The PCR ReadyMix is a Hot Start blend, which prevents amplification of nonspecific products and results in increased efficiency and higher yields of the desired sequence.

Extraction and amplification of genomic DNA from blood occurs in just 3 easy steps.

  1. 10 µl of Extraction Solution is added to 5 µl of whole blood and incubated at room temperature for 5 minutes.
  2. 90 µl of Neutralization Solution is added to the extract.
  3. PCR reactions are setup using 2 µl of the extracts.


Automation Resources

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



Features & Benefits


Rapid Procedure 96 samples of whole blood can be extracted and setup for PCR in 25 minutes. There is no need for organic extraction, centrifugation, column purification, filtration, or alcohol precipitation.
Enhanced Productivity Walk-away automation of genomic DNA extraction and PCR reaction setup allows you to concentrate on other aspects of your work.
High Specificity Specially formulated PCR ReadyMix is a Hot Start blend 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 Blood PCR Kit  
Product No. Product Name Number of Extractions Number of Amplifications
XNAB2R Extract-N-Amp Blood PCR 1000 1000

REDExtract-N-Amp Blood PCR Kit
 
Product No. Product Name Number of Extractions Number of Amplifications
XNABR REDExtract-N-Amp Blood PCR 1000 1000

Kit Contents

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

 



PCR analysis of genomic DNA isolated from 88 blood samples using an automated method for the Extract-N-Amp Blood PCR Kit.
 

Figure 1. Agarose gel analysis of 96 PCR samples.

DNA was extracted from 88 samples of human whole blood (5µl) from a single donor using the automated Extract-N-Amp™ Blood PCR procedure for the Sciclone ALH 3000 Workstation. Amplification of the 236 bp fragment of the HLA gene followed using 2µl of extracted template and 2µl of human genomic DNA controls (A12: 3.2ng/µl, C12: 1.6ng/µl, E12: 0.8ng/µl, G12: 0.2ng/µl) in a 20µl PCR reaction incorporating the 2X PCR Ready Mix. Lane B12, D12, F12, and H12 represent no-template controls. 6µl of each reaction was analyzed on a 1.5% agarose gel.



Cross Contamination Analysis of the Automated Extract-N-Amp Blood Method.

Figure 2. Agarose gel analysis of cross contamination.

5 µl samples of blood or PBS were placed in alternating wells of a microtiter plate. The 96-well plate was processed using the automated Extract-N-Amp Blood method on the Sciclone ALH 3000 Workstation. All samples were then subjected to amplification and 6 µl of the PCR reactions were subjected to electrophoresis on a 2% agarose gel. No PCR products were detected in the wells containing PBS controls.

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