FlexiGene DNA System

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.

Features and benefits

Easy handling - purification in a single tube reduces plasticware consumption and minimizes the risk of sample mix-up
Flexibility in amount of starting material — purification of DNA from 0.1–20 ml whole blood
Direct purification from whole blood — no erythrocyte lysis
Speed - protocol involves only 3 short centrifugation steps and 3 buffers
No organic extraction
High yields of pure DNA

Product

FlexiGene DNA Kit

Principle

The FlexiGene system uses simple centrifugation to isolate highly pure DNA from whole blood, buffy coat, and cultured cells. No organic extraction or other toxic reagents are necessary, and the procedure is faster to perform than other homemade or commercially available methods (see figure "Simple, Rapid Procedure Using the FlexiGene System"). A simple alcohol precipitation is used to recover high yields of pure DNA.

Procedure

The first step is addition of lysis buffer to the samples (see flowchart). 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").

Simple, Rapid Procedure Using the FlexiGene System

Comparison of DNA isolation from twelve 10 ml blood samples, using the FlexiGene DNA Kit or a kit from Supplier G.

High-Quality Genomic DNA

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.

Applications

DNA purified using the FlexiGene system is sized up to 50 kb, with fragments of 30 kb predominating (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").

Efficient Downstream PCR Analysis

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.

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

FlexiGene DNA Kit

For isolation of DNA from whole blood, buffy coat, and cultured cells in a single tube

Format:	Isolation in a single tube, processed by centrifugation
Sample sources:	Whole blood, buffy coat, and cultured cells
Sample size:	0.1-20 ml (blood); 0.1-0.5 ml (buffy coat); 1-2 x 106 cells (cultured cells)
Preparation time:	1 hour
Typical yield:	42 µg DNA from 1 ml whole blood (with normal white blood cell count; 7 x 106 cells/ml blood)
Elution volume:	0.2–1 ml (depending on sample size)

操作手册: FlexiGene DNA Kit Handbook 02/02 (PDF version, 151KB)

Ordering Information
Product	Contents	Cat. No.
FlexiGene DNA Kit (50)	For purification of DNA from 50 ml whole blood: Buffers, QIAGEN Protease	51204

FlexiGene DNA Kit (250)	For purification of DNA from 250 ml whole blood: Buffers, QIAGEN Protease	51206