作业帮有 提取植物线粒体中DNA的具体步骤啊?
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有 提取植物线粒体中DNA的具体步骤啊?
有 提取植物线粒体中DNA的具体步骤啊?
有 提取植物线粒体中DNA的具体步骤啊?
首先用果胶酶和纤维素酶将植物细胞的细胞壁去掉
然后将原生质层放入清水中,让细胞吸胀破裂
再通过离心的方法分离出细胞器,并提取线粒体
将线粒体研磨配成溶液
由于线粒体的组成有水、蛋白质和脂质
将溶液过虑,然后用浓盐法提取并提纯DNA.
首先先用果胶酶和纤维素酶让植物的细胞壁去掉
然后在让植物细胞吸水胀破，让细胞器都出来
再通过密度梯度离心的方法分出细胞器
分出线粒体后，将线粒体磨碎，融入水中，过滤。
最后就是线粒体的DNA了
EXTRACTION OF MITOCHONDRIAL NUCLEIC ACIDS FROM FRACTIONATED MITOCHONDRIA
1. Resuspend the pelleted mitochondria in 0.2 mL of mitochondrial nucleic acids
extraction buffer. Add 4 µL of a 1...
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EXTRACTION OF MITOCHONDRIAL NUCLEIC ACIDS FROM FRACTIONATED MITOCHONDRIA
1. Resuspend the pelleted mitochondria in 0.2 mL of mitochondrial nucleic acids
extraction buffer. Add 4 µL of a 10 mg/mL proteinase K solution and incubate
at room temperature for 5 min.
2. Lyse the mitochondrial membranes by addition of 20 µL of 10% SDS and
incubation at room temperature for 30 min.
3. Extract the nucleic acids with 200 µL of phenol–chloroform–isoamyl alcohol
mixture (25\124\11).
4. Transfer the aqueous phase to a new tube. Add 400 µL of ethanol and place the
tube at –20°C for 20 min. Centrifuge at 13,000gav at 4°C for 30 min. Discard the
supernatant. Add 500 µL of 70% ethanol and centrifuge at 13,000gav at 4°C for
5 min. Discard the supernatant. Dry the precipitate by removing the cap of the
tube, leaving it open at room temperature for 20 min.
5. Dissolve the precipitated nucleic acids in TE (100 µL per 2×105 cells). Store
at 4°C.
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