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1570583814新虫 (初入文坛)
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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
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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. 发自小木虫IOS客户端 |
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