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
Blood 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 0.3 ml whole blood to a 1.5 ml microcentrifute tube, add 0.9 ml 1 x Red Blood Cell Lysis Solution, vortex briefly, and incubate for 2 minutes to lysis the red blood cells.
2. Centrifuge 1 min at 13,000 x g to pellet the white blood cells and discard the supernatant by a pipette (repeat red blood cell lysis if cell pellet is red), leaving 20 ml liquid residue, and resuspend the white cells by pipetting.
3. Add 0.3 ml of 1 x Cell Lysis Solution, resuspend cells by a pipette.
4. Incubate at 55°C for 1 hr to overnight until cell lysate becomes completely clear (disrupted).

Protein precipitation

1. Cool sample to room temperature by placing on ice for 1 min.
2. Add 0.1 ml of Protein Precipitation Solution to the lysate and vortex samples for 20 sec.
3. Place sample into an ice bath for 5 min.
4. Centrifuge at 15,000 x g for 5 min.

DNA Precipitation

1. Pour the supernatant containing DNA into a new 1.5 ml microcentrifute 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 microcentrifute tube containing 0.3 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 airing 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.