For scalable isolation of genomic DNA from whole blood, buffy coat, and cultured cells in a single tube
  • Easy handling, no risk of sample mix-up
  • Scalable purification from 0.1–20 ml whole blood
  • Direct purification from whole blood, buffy coat, and cultured cells
  • Fast procedure
  • No organic extraction

The FlexiGene system uses simple centrifugation to isolate highly pure DNA from whole blood, buffy coat, and cultured cells. The FlexiGene DNA Kit contains buffers and QIAGEN Protease for direct, simple purification in a single tube. For automated solutions using FlexiGene reagents, contact Autogen, Inc.

Product Cat. no. List price:
FlexiGene DNA Kit (250)
For isolation of genomic DNA from whole blood, buffy coat, and cultured cells in a single tube with fast kit preparation
51206
£272.00
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The FlexiGene DNA Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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FlexiGene DNA procedure.|Efficient downstream PCR analysis.|High-quality genomic DNA.|
The first step is addition of lysis buffer to the samples. Cell nuclei and mitochondria are pelleted by centrifugation and resuspended in denaturation buffer containing QIAGEN Protease. Following protein digestion, DNA is precipitated by addition of isopropanol, recovered by centrifugation, washed in 70% ethanol, and dried. DNA is resuspended in hydration buffer and is ready for direct use in downstream assays or storage at –20ºC.
|Variable amounts of DNA template were used to amplify the single-copy Hugl gene and a mitochondrial target (tRNAlys/ATPase). Each sample was analyzed 6 times, and reproducible results were achieved. M: markers.|Genomic DNA isolated from whole blood taken from 4 donors. Starting blood volumes are indicated above the lanes. C: blood treated with sodium citrate; E: blood treated with EDTA; M: markers.|
Performance

DNA purified using the FlexiGene system is sized up to 150 kb (see figure "High-quality genomic DNA"). It performs well in amplification and Southern blotting experiments, and is also suitable for archiving (see figure "Efficient downstream PCR analysis").

DNA is purified from 0.1–20 ml (blood), 0.1–2 ml (buffy coat), or 1–2 x 106 cells (cultured cells) in a single tube, processed by centrifugation. Twelve samples can be processed in 1 hour. Depending on sample size, the rehydration volume is 0.2–1 ml.

DNA yields purified from different sample types using the FlexiGene procedure
Sample  
Volume
DNA yield (µg)
Blood*  
0.3 ml
11–14
Blood*  
2.0 ml
75–90
Blood*  
10.0 ml
 
380–460
Buffy coat  
0.5 ml
 
110–130
HeLa cells  
106 cells
 
25–35

* Whole blood with normal white blood cell counts (7x106 cells/ml blood).

Principle

The FlexiGene DNA system enables isolation of genomic DNA from variable amounts of whole blood, buffy coat, and cultured cells. The simple purification procedure is performed in a single tube, which reduces costs and simplifies handling. The purified DNA performs well in sensitive downstream applications.

Procedure
The first step is addition of lysis buffer to the samples (see flowchart "FlexiGene DNA procedure"). Cell nuclei and mitochondria are pelleted by centrifugation and resuspended in denaturation buffer containing QIAGEN Protease. Following protein digestion, DNA is precipitated by addition of isopropanol, recovered by centrifugation, washed in 70% ethanol, and dried. DNA is resuspended in hydration buffer and is ready for direct use in downstream assays or storage at –20ºC. High DNA yields are obtained from whole blood, buffy coat, and cultured cells (see table "DNA yields purified from different sample types using the FlexiGene procedure").
Applications

DNA purified using the FlexiGene DNA kit performs well downstream applications, including,

PCR
Southern blotting
Archiving

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Kit Handbooks
1
For purification of DNA from human whole blood, buffy coat, cultured cells
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Supplementary Protocols
1
This protocol is designed for purification of DNA from up to 10 ml clotted blood using the FlexiGene DNA Kit.
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Safety Data Sheets
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Download Safety Data Sheets for QIAGEN product components.
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