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
Plasma/serum are lysed to solubilize the DNA contents and proteins are removed by protein precipitation and then precipitate DNA with isopropanol and DNA is resuspended in DNA Hydration Buffer after wash with 70% ethanol to remove the contaminants.

Procedures

Cell Lysis

1. Take 0.5 ml plasma or serum to a 1.5 ml microcentrifuge tube, and add 0.12 ml of 5x Lysis Solution and 5 ml Proteinase K solution (20 mg/ml), vortex for 20 sec.
2. Incubate at 55°C for 1 hr to overnight until cell lysate becomes completely clear.

Protein precipitation

1. Cool sample to room temperature by placing on ice for 1 min.
2. Add 0.25 ml of Protein Precipitation Solution to the lysate.
3. Vortex samples at high speed for 20 sec.
4. Place sample into an ice bath for 5 min.
5. Centrifuge at 15,000 x g for 5 min.

DNA Precipitation

1. Pour the supernatant containing DNA into a new 1.5 ml microcentrifuge tube.
2. Centrifuge at 15,000 x g for 5 min.
3. Pour the supernatant to a new microcentrifuge tube; repeat step 1 and 2 until no pellet is seen.
4. Pour the supernatant containing DNA into a new 1.5 ml microcentrifuge tube containing 0.8 ml 100% isopropanol.
5. Mix the samples by inverting gently 50 times and incubate at room temperature for 5 min.
6. Centrifuge at 13,000 x g for 5 min.
7. Pour off the supernatant and drain the tube briefly on clean absorbent paper. Add 1 ml of 70% ethanol and invert the tube several times to wash the DNA pellet.
8. Centrifuge at 13,000 x g for 5 min, carefully pour off the ethanol and do not lose the DNA pellet.
9. Invert tube and drain the tube on clean absorbent paper, remove the remaining liquid by a pipette, and finally allow air dry for 5 min.

DNA Hydration

1. Add 50 ul DNA Hydration Solution.
2. Resuspend the DNA pellet by a pipette for 5 times and rehydrate the DNA by incubation for 10 min at room temperature.
3. Vortex briefly and pulse spin before use, and store at -20°C.

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.

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.