Description

Features
• Rapid purification of high-quality and ready-to-use DNA.
• No organic extraction.
• Consistent, high yields.
• Complete removal of contaminants and inhibitors for reliable results.

Principle
Bacteria are lysed to solubilize the DNA contents and then proteins are removed by precipitation and DNA-containing supernatant is loaded to DNA binding column and contaminants are then washed away and finally pure DNA is eluted.

Procedures

1. 1. Add 500 µl Buffer GB to DNA binding column with collection tube, centrifuge 13,000 x g for 1 min and discard the flowthrough.
   2. Centrifuge 1-5ml bacterial culture at 13,000 x g for 1 min and discard supernatant. Pipet 250 µl Buffer GS into pellet and suspend bacterial cells and transfer to a 1.5 ml microcentrifuge tube.
   3. Add 250 µl of Buffer GL and mix thoroughly by inverting the tube until solution becomes clear. Cell lysis should be done in 5 min.
   4. Add 350 µl Buffer GP and mix thoroughly by inverting the tube for 4-6 times. Centrifuge at 16, 000 x g for 5 min.
   5. Pipet the supernatant onto the column with collection tube and centrifuge at full speed (20,000 x g) for 1 min. If samples remain in the column, repeat centrifugation to completely remove the remaining samples. Discard the flow-through andbut keep collection tube.
   6. Add 500 µl Buffer GW1 to the column with collection tube and add 500 µl Buffer GW1. Centrifuge at 6000 x g for 1 min. Discard the flow-through and keep the collection tube.
   7. Add 500 µl Buffer GW2. Centrifuge at full speed (20, 000 x g) for 3 min. Discard the flow-through and keep the collection tube.
   8. Centrifuge at full speed for 1 min. Discard the flow-through and collection tube.
   9. Place the mini column in a new 1.5 ml microcentrifuge tube and add 50-100 µl Buffer GE and incubate for 2 min. Centrifuge at 6000 x g for 1 min to elute DNA.

Product Citations

1. Chen J-W et al. (2010) Identification of racehorse and sample contamination by novel 24-plex STR system. Forensic Science International: Genetics 4:158-167. Impact factor: 3.940.
Products used and cited: Genorise Hair and Urine DNA Extraction Kits. Article
2. Chen J-W et al. (2014) Identification of sample donor by 24-plex short tandem repeat in a post-race equine plasma containing dexamethasone. SpringerPlus 3:94.  Article
Products used and cited: Genorise Hair/blood/plasma/urine DNA Extraction kits.