FlexiGene DNA Kit

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

Features

  • 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
FlexiGene DNA Kit (250)

Cat. No. / ID: 51206

For isolation of genomic DNA from whole blood, buffy coat, and cultured cells in a single tube with fast kit preparation
¥46,500
The FlexiGene DNA Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

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.

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).

See figures

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").
See figures

Applications

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

  • PCR
  • Southern blotting
  • Archiving

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR techniques, restriction digest, Southern blotting, sequencing
Sample amount100 µl - 20 ml
FormatScalable
Time per run or per prep25 - 45 minutes (+ DNA rehydration)
Main sample typeWhole blood, buffy coat, cultured cells
TechnologySingle tube precipitation method
ProcessingManual
Yield35 µg / ml whole blood
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinGenomic DNA
Elution volume200 µl - 1000 µl

Resources

Kit Handbooks (2)
For purification of DNA from human whole blood, buffy coat, cultured cells
Supplementary Protocols (1)
This protocol is designed for purification of DNA from up to 10 ml clotted blood using the FlexiGene DNA Kit.
Additional Resources (1)
7 years storage at 2-8°C and -20°C

Publications

The (TAAAA)n microsatellite polymorphism in the SHBG gene influences serum SHBG levels in women with polycystic ovary syndrome.
Ferk P; Teran N; Gersak K;
Hum Reprod; 2006; 22 (4):1031-6 2006 Dec 22 PMID:17189294
A functional SNP of interferon-gamma gene is important for interferon-alpha-induced and spontaneous recovery from hepatitis C virus infection.
Huang Y; Yang H; Borg BB; Su X; Rhodes SL; Yang K; Tong X; Tang G; Howell CD; Rosen HR; Thio CL; Thomas DL; Alter HJ; Sapp RK; Liang TJ;
Proc Natl Acad Sci U S A; 2007; 104 (3):985-90 2007 Jan 10 PMID:17215375
Lipopolysaccharide-activated CD4+CD25+ T regulatory cells inhibit neutrophil function and promote their apoptosis and death.
Lewkowicz P; Lewkowicz N; Sasiak A; Tchórzewski H;
J Immunol; 2006; 177 (10):7155-63 2006 Nov 15 PMID:17082633
Predictors of vitamin B6 and folate concentrations in older persons: the InCHIANTI study.
Gori AM; Sofi F; Corsi AM; Gazzini A; Sestini I; Lauretani F; Bandinelli S; Gensini GF; Ferrucci L; Abbate R;
Clin Chem; 2006; 52 (7):1318-24 2006 May 11 PMID:16690736
Past adult lead exposure is linked to neurodegeneration measured by brain MRI.
Stewart WF; Schwartz BS; Davatzikos C; Shen D; Liu D; Wu X; Todd AC; Shi W; Bassett S; Youssem D;
Neurology; 2006; 66 (10):1476-84 2006 May 23 PMID:16717205

FAQ

Do you have a protocol for purification of DNA from clotted blood using the FlexiGene DNA Kit?

Yes, please follow the User-Developed Protocol 'Purification of DNA from clotted blood using the FlexiGene DNA Kit' (FG01).

 

FAQ ID -973
Do you have stability data for genomic DNA isolated with the Flexigene DNA Kit?

Yes. We have shown that DNA purified with the Flexigene DNA Kit is stable for at least 7 years when being stored at either 2-8°C or -20°C.  However, the DNA stability is dependent upon storage conditions.  The data is available here under Additional Resources.

 

FAQ ID -1317
Do you have a protocol for the isolation of genomic DNA from frozen clotted blood?

Yes, we have the following protocols:

  • Isolation of genomic DNA from frozen clotted whole blood using the QIAGEN Genomic-tip 100/G (QG02). TEST

You will need to prepare the required buffers according to the recipes in Appendix A of the QIAGEN Genomic DNA Handbook, or you can purchase the Genomic DNA Buffer Set containing pre-made solutions. Alternatively, the QIAGEN Blood & Cell Culture Midi Kit, containing Genomic-tips 100/G and buffers, can be used.

  • Purification of archive-quality DNA from clotted whole blood using Clotspin Baskets and the Gentra Puregene Blood Kit (PG03).
  • Purification of archive-quality DNA from clotted whole blood using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit (PG04).
  • Purification of DNA from clotted blood using the FlexiGene DNA Kit (FG01).
FAQ ID -900. Test
What is the composition of Buffer FG3?
Buffer FG3, the hydration buffer supplied in the FlexiGene DNA Kit, is a 10 mM Tris-Cl solution at pH 8.5.
FAQ ID -356