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1570583814新蟲 (初入文壇)
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[求助]
求蛋白質(zhì)提取方法的簡潔順暢英文概括，具體步驟如下 已有1人參與
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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. 發(fā)自小木蟲IOS客戶端 |
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★ ★
感謝參與，應助指數(shù) +1
空位: 金幣+2, 金幣獎勵，辛苦了o(∩_∩)o 2016-03-18 14:15:08
感謝參與，應助指數(shù) +1
空位: 金幣+2, 金幣獎勵，辛苦了o(∩_∩)o 2016-03-18 14:15:08
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2樓2016-03-09 10:23:45
1570583814
新蟲 (初入文壇)
- 應助: 0 (幼兒園)
- 金幣: 315.9
- 散金: 70
- 帖子: 40
- 在線: 14.9小時
- 蟲號: 4170639
- 注冊: 2015-10-24
- 性別: MM
- 專業(yè): 作物種質(zhì)資源與遺傳育種學
3樓2016-03-09 13:10:39
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