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【有奖交流】积极回复本帖子，参与交流，就有机会分得作者 1570583814 的 30 个金币

1570583814

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应助: 0 (幼儿园)
金币: 315.9
散金: 70
帖子: 40
在线: 14.9小时
虫号: 4170639
注册: 2015-10-24
性别: MM
专业: 作物种质资源与遗传育种学

[交流] 求以下蛋白质提取方法的简洁连贯英文版，具体操作步骤英文叙述如下

初始悬赏金币 30 个
1) The sample was grinded to power with liquid nitrogen;
2) Add 10mL cooled acetone contained 10% TCA to 1g sample power at  -20°C for 1 hour;
3) Centrifuge by 15000g for 15 min at  4°C, collect the  deposit  and  add cooled  acetone at -20°C for 1 hour;
4) Repeated the step 3;
5) Centrifuge  by  15000g  for  15  min  at  4°C,  collect  the  deposit  and  dried  by vacuum freezed dryer;
6) Suspends the deposit in cold phenol extraction buffer;
7) Add an equal volume of  phenol saturated with Tris-HCl  (pH  7.5) and shake  the mixture for 30 min at 4 ℃;
8) Centrifuge at 5000g for 30 min at 4 ℃ and collect the upper phenolic phase;
9) Add  equal  phenol  extraction  buffer  to  the  collected  phenolic  phase,  repeat steps 2-3 and collect the upper phenolic phase again;
10) Add 5 volumes of cold 0.1 M ammonium acetate in methanol to the collected phenol phase, store at –20 ℃ for 1 hour;
11) Centrifuge the sample for 30 min at 5000g at 4 ℃;
12) Add 2 volumes (based on the volume of the last collected phenolic phase) ice-cold methanol to wash the pellet, mix gently;
13) Centrifuge the sample for 30 min at 5000g at 4 ℃;
14) Repeat Steps 12and 13 two more times;
15) Repeat Steps 12 and 13 two more times using acetone instead of methanol;
16) Centrifuge the sample for 30 min at 5000g at 4 ℃, dry the deposit;
17) The deposit was dissolved in lysis solution at 30°C for 1 hour;
18) Centrifuge the solution by 15000g for 15 min at room temperature, collect the supernatant and centrifuge again;
19) The supernatant was the extracted protein solution.

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