碱裂解法提取质粒为什么加溶液ⅱ裂解的时间不能过长

对牛弹琴冲淡尚 2015-04-24 02:54:04 420 浏览

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顾天虹123 2015-04-25 00:00:00

哦

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庞小芹 2015-04-30 00:00:00

你多放上一会，NaOH就把基因组DNA全弄碎了，跟质粒混在一起分不开了

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热门问答

碱裂解法提取质粒为什么加溶液ⅱ裂解的时间不能过长:

碱裂解法小量提取质粒，加TE后，为什么不溶解？:

碱裂解法提取质粒中为什么要冰浴:

结合碱裂解法提取质粒，配置溶液1为什么使用ph酸度计？:

结合碱裂解法提取质粒，配置溶液1为什么使用ph酸度计:

碱裂解法提取质粒过程溶菌酶的作用:

碱裂解法提取质粒dna的注意事项有哪些:

SDS碱裂解法提取质粒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！哪位专家能帮帮忙啊？... 想知道溶液1的成分及其作用！溶液2，溶液3也想知道！哪位专家能帮帮忙啊？展开

碱裂解法提取质粒时，先加的缓冲液使菌体悬浮的原因是什么:

碱裂解法提取的质粒DNA琼脂糖凝胶电泳应该是什么样的?:

碱裂解法提取质粒DNA时的SDS有什么作用:

碱裂解法提取质粒后电泳拖尾严重，是什么原因:

翻译protocol（有关质粒DNA提取，碱裂解法）？: 请哪位高人帮忙找道这篇的中文：PreparationofPlasmidDNAbyAlkalineLysiswithSDS:Minipreparation（题）或者帮忙翻译一下，要求不要用电脑直接翻译，总之通顺准确即可。急需截止25号。... 请哪位高人帮忙找道这篇的中文：Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation（题）或者帮忙翻译一下，要求不要用电脑直接翻译，总之通顺准确即可。急需截止25号。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. 展开

碱裂法提取质粒加完溶液二为什么变澄清:

碱裂解法提取质粒中的乙酸钾可以换成乙酸钠吗:

碱裂解提取质粒DNA可以酶切吗:

有些微生物培养基恒温培养时间为什么不能过长？谢谢:

碱裂解法提取质粒主要使用哪些试剂？这些试剂的主要作用是什么:

碱裂解法提取质粒DNA的电泳图谱，为何我的只有两条带？分别都是什么？: 1.我的Marker选的100bp是不是不合适？2.这两条带是线性和开环构型DNA？？这两条带哪个是线性哪个开环构型DNA？超螺旋的为什么没有？... 1.我的Marker 选的100bp 是不是不合适？ 2.这两条带是线性和开环构型DNA？？这两条带哪个是线性哪个开环构型DNA？超螺旋的为什么没有？展开

用碱裂解的方法提取细菌基因组DNA的具体方法是什么？不是提取质粒。: 因为用赛百盛公司的基因组DNA提取试剂盒效果不是很好，准备换用碱裂解的方法提取。但是碱裂解的方法查到的资料都是提取质粒DNA的，没有提取基因组DNA的方法。大家有知道的麻烦告诉我一... 因为用赛百盛公司的基因组DNA提取试剂盒效果不是很好，准备换用碱裂解的方法提取。但是碱裂解的方法查到的资料都是提取质粒DNA的，没有提取基因组DNA的方法。大家有知道的麻烦告诉我一声，谢谢！另外，大家都是用那种提取基因组DNA的试剂盒提取啊？效果那些比较好？谢谢大家了！展开

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