BP-10 Spin Column DNA Gel Extraction minipreps kit
Components | DGE-50, 50 Preps | DGE-100, 100 Preps |
Binding Buffer II Wash Solution Elution Buffer BP-10 Column 2.0ml Collection tube | 1X30ml 12ml 5ml 50 50 | 2X30ml 24ml 10ml 100 100 |
(A)Before use, add 48ml of 96-100% of ethanol to 12ml
(B)Elution Buffer is 2mM Tris-HCl pH 8.0~8.5. Although TE buffer pH 8.0 or water may be substituted, the resulting yields may be up to 20% lower.
Storage: The kit is stable for 12 months at room temperature. For longer storage, keep all contents of the kit cold.
Principle:
The BP-10 spin column purification kit utilizes a silica-gel based membrane which selectively adsorbs up to 10ug of DNA fragments in the presence of specialized binding buffers. Nucleotides, oligos (<40-mer), enzymes, mineral oil and other impurities do not bind to the membrane and are washed away.DNA fragments are then eluted off the column and can be used for downstream protocols withoutfurther processing.
Application:
- Recovery of DNA fragments from reaction solutions.
- Recovery of DNA fragments from agarose gels.
Features:
- Quick and economical
- High yields (60-90%) of 60bp-40kb DNA fragments.
- Efficient removal of contaminants. Purified DNA can be used in any downstream application such as sequencing, labeling, restriction enzymatic digestion, ligation or transformation.
- No phenol / chloroform extraction or ethanol precipitation is required.
Protocol for Agarose Gel:
1. Excise the DNA fragment from the gel with a clean, sharp scalpel. Weigh the gel slice and transfer to a 1.5mL microfuge tube.
2. Add 400ul of Binding Buffer II for each 100mg of gel weight (example - a gel slice weighing 125mg would require 500ul of Binding Buffer II). Incubate at 50-600C for 10 minutes and shake occasionally until agarose is completely dissolved. For high concentration gels (1.5-2.0%), 700ul of Binding Buffer II per 100mg of agarose gel are added.
3. Add the above mixture to the BP-10 column and let stand for 2 minutes.Centrifuge at 8,000 x g for 1 minute and discard the flow-through in the tube.
4.Add 500ul of Wash Solution, and centrifuge at 8,000 x g for one minute. Discard the solution in the tube.
5.Repeat step 5. Centrifuge at 12,000 x g for an additional 1 minute to remove any residual Wash Buffer.
6.Place the column in a clean 1.5ml microfuge tube.Add 30-50ul of Elution Buffer to the center of the column and incubate at room temperature for 2 minutes. Centrifuge at 12,000 x g for 1 minute to elute DNA.
Note: It is extremely important to add the Elution Buffer to the center of the column. Incubating the column at higher temperatures (37º to 50ºC) may slightly increase the yield. Or prewarming the Elution Buffer at 55ºC to 80ºC may also slightly increase elution efficiency.
If a higher DNA concentration is desirable, 20ul (or less) of elution buffer can be used to elute the DNA. It is critical that the elution buffer be applied directly in the center of the column. (To recover maximum amount of DNA it is recommended to repeat the elution step.)
7.Store the purified DNA at -20ºC.
Protocol for DNA purification from enzymatic reactions:
1.Transfer the volume of the reaction mixture to a 1.5ml microfuge tube and add 3 volumes of Binding Buffer II. Mix by inverting the tube a few times.
2.Add the above mixture to the BP-10 column and let stand for 2 minutes. Centrifuge at 5000rpm for 1 minute and discard the flow-through in the tube.
3.Add 500ul of Wash Solution, and centrifuge at 8,000rmp for 1 minute. Discard the solution in the tube.
4.Repeat step 3. Spin at 1000rpm for an additional minute to remove any residual
5.Place the column in a clean 1.5ml microfuge tube. Add 30-50ul of Elution Buffer to the center of the column and incubate at room temperature for 2 minutes.
Note:It is extremely important to add the Elution Buffer to the center of the column.Incubating the column with the Elution Buffer at higher temperatures may slightly increase the yield, especially of fragments larger than 10,000bp. Prewarming the Elution Buffer at 55º to 80ºC may also slightly increase elution efficiency. If a higher DNA concentration is desirable, 20ul (or less) of elution buffer may be used. It is recommended that the elution step be repeated to recover the maximum amount of DNA.
6.Centrifuge at 1000rpm for 1 minute to elute the DNA.
7.Store the purified DNA at -20ºC.
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求助各位前辈，我最近在合成的化合物水溶性很好，非常好，以至于可以随便溶解在水里，它的六氟磷酸盐也可以随意溶解在水里（大于50uM），细胞成像实验显示它根本进不去细胞，求问有没有啥方法包裹一下让它进去？我搜了一下文献，感觉多数是把脂溶性特别好的东西包裹一下弄进去的，也许是搜索姿势不对没找到我需要的答案，**点拨啊！！！
求助各位大神，现在想购买小分子数据库，求大神推荐。
我知道的免费的数据库有zinc
求推荐哪家公司或者研究所的小分子数据库可以购买，十分感谢！！！！！！
求助大家，小分子药物最新专利申请情况跟踪，之前听别人说可以在一个网站可以导出这个信息。
不知哪位大侠知道，告知一下，不胜感激！
就是蛋白质分子的小片断
是氨基酸形成的
一、首先你要明白肽是什么...........................肽是氨基酸通过酰胺键结合而成的东西.........
二、你要明白氨基酸是什么..........................氨基酸是构成蛋白质的基本单位，多种氨基酸结合为长肽链，几条长肽链再盘旋就形成了蛋白质......................
三、关于小分子肽、短肽、多肽、寡肽.......其实都是肽.......区别只是由多少个氨基酸构成而已............
所以，你的问题可以很粗暴地理解为“蛋白质对人体有没有副作用”.......
如果你营养足够的情况下，再补充这个，会导致营养过剩，从而加重身体代谢的负荷............类似就是这样子的了...........小分子肽，一般现在用于化妆品上比较多（一ye子.植物肽面膜就是这个）.......小分子肽（可以简单理解为纳米胶原蛋白），这比蛋白质（也可以粗暴理解为胶原蛋白）更加容易吸收......而用在食品上，要视乎是何种小分子肽了.....大豆肽、花生肽、大米肽....不同的肽有不同的功效.......主要可以改善风味、改善吸收、增强胃肠道功能等等.......
有机的是有机化合物的简称,它指的是含碳化合物.
但是,有四大类常见物质一般不作为有机物处理：
1、碳的氧化物,如CO和CO2.
2、碳酸及其盐,如CaCO3.
3、金属碳化物,如CaC2.
4、拟卤素及其化合物,如(CN)2与KSCN.
水的化学式为H2O,它不含有碳元素,故不是有机物.
但若所描述的水不是化学意义的水,而是自然界存在的天然水,那么,水中会溶有一定量的有机物.
