結合堿裂解法提取質粒，配置溶液1為什么使用ph酸度計？

sunyuyuan10 2018-03-24 16:49:52 393 瀏覽

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yangle406 2018-03-25 00:00:00

需要控制酸度

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結合堿裂解法提取質粒，配置溶液1為什么使用ph酸度計:

結合堿裂解法提取質粒，配置溶液1為什么使用ph酸度計？:

堿裂解法提取質粒中為什么要冰浴:

堿裂解法提取質粒為什么加溶液ⅱ裂解的時間不能過長:

SDS堿裂解法提取質粒DNA中溶液1的成分及作用是什么？: 想知道溶液1的成分及其作用！溶液2，溶液3也想知道！哪位專家能幫幫忙啊？... 想知道溶液1的成分及其作用！溶液2，溶液3也想知道！哪位專家能幫幫忙啊？展開

堿裂解法提取質粒過程溶菌酶的作用:

堿裂解法小量提取質粒，加TE后，為什么不溶解？:

堿裂解法提取質粒dna的注意事項有哪些:

堿裂解法提取質粒主要使用哪些試劑？這些試劑的主要作用是什么:

堿裂解法提取質粒后電泳拖尾嚴重，是什么原因:

堿裂解法提取質粒DNA時的SDS有什么作用:

堿裂解法提取的質粒DNA瓊脂糖凝膠電泳應該是什么樣的?:

如何使用酸度計測量溶液的ph:

翻譯protocol（有關質粒DNA提取，堿裂解法）？: 請哪位高人幫忙找道這篇的中文：PreparationofPlasmidDNAbyAlkalineLysiswithSDS:Minipreparation（題）或者幫忙翻譯一下，要求不要用電腦直接翻譯，總之通順準確即可。急需截止25號。... 請哪位高人幫忙找道這篇的中文：Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation（題）或者幫忙翻譯一下，要求不要用電腦直接翻譯，總之通順準確即可。急需截止25號。原文如下： 1. Inoculate 2 ml of rich medium (LB， YT， or Terrific Broth) containing the appropriate antibiotic with a single colony of transformed bacteria. Incubate the culture overnight at 37°C with vigorous shaking. 2. Pour 1.5 ml of the culture into a microfuge tube. Centrifuge at maximum speed for 30 seconds at 4°C in a microfuge. Store the unused portion of the original culture at 4°C. 3. Remove the medium by aspiration， leaving the bacterial pellet as dry as possible. 4. Resuspend the bacterial pellet in 100 μl of ice-cold Alkaline lysis solution I by vigorous vortexing. 5. Add 200 μl of freshly prepared Alkaline lysis solution II to each bacterial suspension. Close the tube tightly， and mix the contents by inverting the tube rapidly five times. Do not vortex! Store the tube on ice. 6. Add 150 μl of ice-cold Alkaline lysis solution III. Close the tube and disperse Alkaline lysis solution III through the viscous bacterial lysate by inverting the tube several times. Store the tube on ice for 3-5 minutes. 7. Centrifuge the bacterial lysate at maximum speed for 5 minutes at 4°C in a microfuge. Transfer the supernatant to a fresh tube. 8. (Optional) Add an equal volume of phenol:chloroform. Mix the organic and aqueous phases by vortexing and then centrifuge the emulsion at maximum speed for 2 minutes at 4°C in a microfuge. Transfer the aqueous upper layer to a fresh tube. 9. Precipitate nucleic acids from the supernatant by adding 2 volumes of ethanol at room temperature. Mix the solution by vortexing and then allow the mixture to stand for 2 minutes at room temperature. 10. Collect the precipitated nucleic acids by centrifugation at maximum speed for 5 minutes at 4°C in a microfuge. 11. Remove the supernatant by gentle aspiration as described in Step 3 above. Stand the tube in an inverted position on a paper towel to allow all of the fluid to drain away. Use a Kimwipe or disposable pipette tip to remove any drops of fluid adhering to the walls of the tube. 12. Add 1 ml of 70% ethanol to the pellet and invert the closed tube several times. Recover the DNA by centrifugation at maximum speed for 2 minutes at 4°C in a microfuge. 13. Remove all of the supernatant by gentle aspiration as described in Step 3.Take care with this step， as the pellet sometimes does not adhere tightly to the tube. 14. Remove any beads of ethanol that form on the sides of the tube. Store the open tube at room temperature until the ethanol has evaporated and no fluid is visible in the tube (5-10 minutes). 15. Dissolve the nucleic acids in 50 μl of TE (pH 8.0) containing 20 μg/ml DNase-free RNase A (pancreatic RNase). Vortex the solution gently for a few seconds. Store the DNA solution at -20°C. 展開