Extract-N-Amp™ Blood Product Information

Description

The Extract-N-Amp Blood PCR kits contain all of the reagents necessary to rapidly extract genomic DNA from whole blood 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 Blood PCR Kit offers a novel extraction system that eliminates the need for DNA purification, organic extraction, centrifugation, heating, filtration or alcohol precipitation. 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.

Procedure

Genomic DNA is extracted from 10 µl of whole blood by adding Extraction Solution and incubating for 5 minutes at room temperature. After adding Neutralization Solution, this crude blood extract is stable for at least 6 months at 4 °C and 3 months at 37 °C (Figure 2) and can be added directly to the optimized PCR mix supplied.

xnab-flow-chart

Application

Perfect for cloning and automated sequencing protocols.

Beneficial Features

 

Starting Material 10 µl of blood
Speed Extraction of genomic DNA for PCR in 8 minutes
Scalable Liquid format fits any configuration (single tube, 96-well, etc.) (Figure 1)
Specific Hot Start antibody for highly specific PCR amplification of genomic DNA
Safe No phenol/chloroform or other hazardous materials

 

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.

Ordering Information

Extract-N-Amp Blood PCR Kits

Product No. Name Number of Extractions Number of Amplifications Technical Manual .pdf
XNAB2 Extract-N-Amp Blood PCR Kit 100 100 X
XNAB2R Extract-N-Amp Blood PCR Kit 1000 1000 X
XNAB2RE Extract-N-Amp Blood PCR Kit 1000 5000 X

 

REDExtract-N-Amp Blood PCR Kits

Product No. Name Number of Extractions Number of Amplifications Technical Manual .pdf
XNABS REDExtract-N-Amp Blood PCR Kit
(contains REDTaq)
10 10 X
XNAB REDExtract-N-Amp Blood PCR Kit
(contains REDTaq)
100 100 X
XNABE REDExtract-N-Amp Blood PCR Kit
(contains REDTaq)
100 500 X
XNABR REDExtract-N-Amp Blood PCR Kit
(contains REDTaq)
1000 1000 X
XNABRE REDExtract-N-Amp Blood PCR Kit
(contains REDTaq)
1000 5000 X

 

Kit Contents

Extract-N-Amp Blood PCR Kit

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

Sample Data

PCR analysis of genomic DNA isolated from blood using Sigma's Extract-N-Amp Blood PCR Kit

PCR analysis of genomic DNA isolated

Figure 1. Extract-N-Amp Blood PCR Kit used to isolate genomic DNA from fresh, 7 day old & frozen blood. DNA was extracted and neutralized from 10 µl of whole blood in 5 minutes at room temperature using the REDExtract-N-Amp Blood PCR kit. The PCR products were then generated using the specially formulated Hot Start PCR mix included in the kit. PCR products generated are 1.8 kb for carnitine palmitoyltransferase II, 1.3 kb for a mitochondrial DNA control region, 547 bp for human surfactant protein B and 320 bp for the 5' untranslated region of human major histocompatibility complex class II.

Stability of Extract-N-Amp Blood Extracts

Stability of Extract-N-Amp Blood

Figure 2. Blood was drawn from 2 human volunteers into vacutainer tubes containing EDTA. Extractions were performed in duplicate providing 4 samples total. Samples were stored at 37 °C and removed at various time intervals for testing. Stability was determined by monitoring yield from quanitative PCR using an ABI 7700 instrument. The DNA standards used for the quanitative PCR were generated from the same blood drawn as the test samples. The DNA for the standards was purified using the GenElute Blood Genomic DNA Kit (NA2000). The PCR products were generated using primers for a 547 bp product for human surfactant protein B (SPB; Lin & Floros, 2000, BioTechniques, 29:460-466). The results clearly show no loss of amplification of the SPB PCR product even after storage at 37 °C for 6 months.

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