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6 years ago in Discussion By Yashti Iyer

Techniques to isolate DNA from the blood

I want to isolate DNA from the blood. Which all techniques can I use for this purpose and how do I do it

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By Zeba Malik Answered 6 years ago

The best method that can be used here is Salt-Out Method.
The fresh/frozen blood, after thawing, should be resuspended in nine volume of Buffer A 10 mL 1 M Tris HCl (pH 7.6), (109.5 g sucrose , 1% Triton-X-100, 5 mL 1 M MgCl2 ,) and must be kept on ice for 10- 15 min. This is then followed by 10 min centrifuge (10,000 rpm at 4 o C). The supernatant must be discarded and nuclei pellets should be resuspended in 1.8 mL of Buffer B (75 mM NaCl, 20 µg/mL pancreatic Ribonuclease A, 25 mM EDTA (pH 8.0),) and 150 µl of 20 µl Proteinase K (20 mg/mL and 10% SDS). The suspension must then be incubated overnight (56 o C) on low speed rocker/orbiter shaker in water bath. Saturated NaCl must be added and the sample must be vigorously mixed for around 15 min. This is then followed by spinning at 10,000 rpm (at 4 o C for 10 min). The supernatant should be then transferred to another 15 mL polypropylene tube, and also the pellets should be discarded. The DNA must be precipitated by mixing supernatant and you will obtain the desired result.

By Kamlesh Meena Answered 6 years ago

Yashti, you can use salt-out-method or you can also use, Phenol – Chloroform Method. In this method fresh and frozen blood samples must be treated with RBC lysis buffer only after thawing. After centrifugationWBC pellets must be suspended in extraction buffer. DNA threads should then be washed with 70% ethanol.

By Muhammad Umar Farooq Answered 6 years ago

Yashti, kindly go though https://www.sciencedirect.com/science/article/pii/S0344033806000628. This site consists of a research paper based on the DNA extraction. It might help you.

By Yashti Iyer Answered 6 years ago

Thank you so much for the suggestions.

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