计算溶液所需的质量、体积或浓度。
BioReagent,DNA级,分子生物学级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
2-8°C储存,充氩。低温运输 。请查阅批次 COA 获取详细规格。
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在色谱分析、有机合成和交叉偶联反应领域已被 3 篇同行评审文献引用。
Sephadex™ G-100 DNA级色谱树脂用于从小分子(如盐)中通过尺寸排阻法纯化长度至多25个碱基的DNA片段。与较小分子相比,长度超过25个碱基的DNA片段将被树脂排除,不会保留而直接通过。该介质以散装(干燥)形式提供,以便研究人员自行制备用于纯化核酸的色谱柱。该产品经过测试,确保其可重复性和DNA的高回收率(>90%),非常适合通过凝胶过滤进行高回收率的核酸纯化。
使用说明
葡聚糖凝胶粉末在使用前必须进行水合。在装入柱子之前,树脂必须充分溶胀。
溶胀时间:20°C下72小时或90°C下5小时
第1步:称取所需床体积的适量干凝胶(床体积约为:15–20 mL/g干凝胶)。
第2步:加入足够的缓冲液,使其总量等于色谱柱的总体积,并额外多加30%。
第3步:搅拌。应避免过度搅拌,以免破坏珠粒:切勿使用磁力搅拌器。使用沸水浴可加速该过程,同时也有助于缓冲液脱气。
第4步:倾析上清液(完全溶胀后)以去除细小颗粒。 第5步:重新加入缓冲液制成悬浮液;此时悬浮液应相当浓稠(约75%为沉淀凝胶)。
第6步:如需脱气,建议使用连接真空线的合适玻璃容器。准备完毕后,倒入合适的色谱柱中。为达到平衡,需使待分离的水性缓冲液通过色谱柱2至3倍柱体积。溶胀树脂可在2°C–8°C下冷藏保存,最长可达1个月。若需长期保存,应在缓冲液中加入抑菌剂,如20%乙醇或Kathon。
洗脱液组成不应直接影响可获得的分辨率:1. 对于携带带电基团的物质,使用含有缓冲液(如Tris-HCl、磷酸钠)的洗脱液来控制pH值,并建议离子强度至少为0.02,以防止与凝胶基质可能发生的离子相互作用。NaCl可用于此目的。 2. 对于即将进行冻干的产品:使用挥发性缓冲液,如乙酸铵、碳酸氢铵或乙二胺乙酸盐。
Sephadex™ G-100 DNA Grade chromatography resin for purification of DNA fragments up to 25 bases in length from small molecules such as salts by size exclusion. DNA fragments above 25 bases will be excluded from the resin and will pass through without retention compared to smaller molecules. The media is provided in a bulk (dry) format to allow researchers to prepare their own columns for purifying nucleic acids.The product is tested to ensure reproducibility and high recovery of DNA (> 90%) and is highly suitable for high-recovery nucleic acid purification via gel filtration.
Instructions for use
Sephadex powder has to be hydrated before use. The resin must be fully swollen before packing into columns.
Swelling time: 72 hours at 20°C or 5 hours at 90°C
Step 1
Weigh out the appropriate amount of dry gel for the required bed volume (approx. bed volume: 15–20 mL/g dry gel).
Step 2
Step 2 Add enough buffer to equal the total volume of the column plus 30% more.
Step 3
Stir. Excessive stirring should be avoided as it may break the beads: DO NOT USE MAGNETIC STIRRERS. The process is accelerated by using a boiling water bath, which also serves to deaerate the buffer.
Step 4
Decant the supernatant (after complete swelling) to remove fines.
Step 5
Reintroduce buffer to make a suspension; this should be fairly thick (about 75% settled gel).
Step 6
If degassing is required, use of an appropriate glass vessel attached to a vacuum line is recommended.Once prepared, pour into an appropriate column. To equilibrate, pass 2 to 3 column volumes of the aqueous buffer to be used in the separation.Swollen resin can be stored refrigerated at 2°C–8°C. for up to 1 month. For longer term storage, a bacteriostatic agent should be included in the buffer such as 20% ethanol or Kathon.
Eluent composition should not directly influence the resolution that can be obtained:
1. For substances carrying charged groups an eluent containing a buffer e.g. Tris-HCl, sodium phosphate is used to control pH, and an ionic strength of at least 0.02 is recommended to safeguard against possible ionic interactions with the gel matrix. NaCl can be used forthis purpose.
2. For product that is to be lyophilized: use volatile buffers e.g. ammonium acetate, ammonium bicarbonate or ethylenediamine acetate.
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