FlexiGene DNA Kit

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

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

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

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.

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").

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

• PCR
• Southern blotting
• Archiving