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The Easiest Route to Guaranteed SilencingIncreasing numbers of researchers are using small interfering RNAs (siRNAs) to reduce the expression of specific mammalian genes. Applications of siRNA induced ...

Restriction Map and Cloning Site of the RNAi-Ready pSIREN-DNR Vector. Unique restriction sites are in bold. RNAi-Ready pSIREN-DNR is provided as a linearized vector digested with BamH I and EcoR I. N ...

RNA Marker ( RL1000; RL6000; RL10000) 可以进行一般琼脂糖凝胶电泳和变性琼脂糖凝胶电泳。以RL10000为例，具体使用方法如下：普通琼脂糖凝胶电泳时1.按下列组份配置RNA Marker样品。2.均匀混合后65℃加热10分钟，迅速冷却至室温（最好用PCR仪)。3.使用高质量的琼脂糖，用1×TAE Buffer制备3%凝胶，制胶时凝胶中请加入溴乙锭(最终浓度：1 ...

在人细胞中快速、高水平地表达目的蛋白 在2至3个星期内即可获得高效价的病毒 无需噬斑纯化 靶细胞可以是分裂或不分裂的细胞（dividing/or nondividing cell） CLONTECH的Adeno-X™表达系统是一个特别 高效的腺病毒表达系统，用于在人细胞中瞬时及高水平地表达目的蛋白质。由于Adeno-X™病毒DNA中含有特别设计的克隆位点，你可以在2星期内获得高效价的腺病毒 ...

1．分别设立两个反应，用适当的限制性内切核酸酶消化1~10μg质粒DNA和外源DNA片段，使它们能产生平末端。2．苯酚：氯仿抽提与乙醇沉淀法纯化出已被消化的载体和外源DNA片段。3．分别用TE（pH 8.0）重新溶解纯化出的两种DNA沉淀，使终浓度为100 ng/ml。 假设1 bp相当于660 Da，计算DNA的终浓度（pmol/ml）。4．将载体DNA去磷酸化。5．按下表将适量的DNA转移至无 ...

体外定点突变技术是研究蛋白质结构和功能之间的复杂关系的有力工具，也是我们在实验室中改造/优化基因常用的手段。蛋白质的结构决定其功能，二者之间的关系是蛋白质组研究的重点之一。对某个已知基因的特定碱基进行定点改变、缺失或 者插入，可以改变对应的氨基酸序列和蛋白质结构，对突变基因的表达产物进行研究有助于我们了解蛋白质结构和功能的关系，探讨蛋白质的结构/结构域。而利用定点突变技术改造基因，相信 ...

TRTn3 terminal inverted repeatslacZ5'-truncated lacZ gene encoding beta-galactosidaseLEU2LEU2 gene from S. cerevisiaeampEncodes beta-lactamaseloxPlox site target for Cre recombinaseUses: Gene disrupti ...

TRTn3 terminal inverted repeatsHAHemagglutinin (HA) epitope loxRlox site target for Cre recombinaselacZ5'-truncated lacZ gene encoding beta-galactosidaseURA3URA3 gene from S. cerevisiaetetTetracycline ...

Identify appropriate celltype over-expressing corresponding gene.Find out if transcription can be stimulated further eg by induction (note: the higher the mRNA level of the gene in question the easier ...

Plasmid isolated by this procedure can be used routinely for electrophoretic analysis restriction endonuclease digestion and transformation of E. Coli. sequencing PCR and most other molecular biologic ...

The gel shift or electrophoretic mobility shift assay provides a simple and rapid method for detecting DNA-binding proteins. This method has been used widely in the study of sequence-specific DNA-bind ...

1) The ligation mixture contains the following: vector DNA (~100 ng) insert DNA (equimolar or 2 or 3 X molar concentration of vector) water added to 18 µl 2) Heat the mixture at 45 °C for 5 min. to me ...

Prepare a ligation mix:Ligation Mix (2x)10x ligase buffer1.0 mldigested vector(0.1 mg/ml)1.0 mlH2O6.0 mltotal8.0 mldivide ligation mix into two Eppendorf tubesLigation Rxn InsertControlligation mix ...

Prepare a 100 µl reaction mixture containing the DNA of interest in the appropriate 1X restriction enzyme buffer. Divide the reaction mixture between five prechilled microcentrifuge tubes such that tu ...

Gel Shift or Band Shift Assay or Electrophoretic Mobility Shift Assay (EMSA) is a technique for studying gene regulation and determining protein:DNA interactions. The assay is based on the observation ...

Q：利用凝胶回收试剂盒DNA回收效率较低或者未回收到目的片段？A：可能有以下几个原因： 1. 胶块未完全溶解，可适当延长水浴时间，并增加上下颠倒次数辅助溶解； 2. 胶块体积太大，应先将其切为小块，分多次回收； 3. 电泳缓冲液pH太高，硅基质膜在高盐低pH结合DNA，如pH太高，应作适当调整；4. 漂洗液中未加入无水乙醇。Q：能否改用去离子水洗脱？A：可以。但是实验室所用纯水一般 ...

　　蛋白酶k是一种枯草蛋白酶类的高活性蛋白酶，用于生物样品中蛋白质的一般降解。从林伯氏白色念球菌（tritirachium album limber）中纯化得到。该酶有两个ca2+结合位点，它们离酶的活性中心有一定距离，与催化机理并无直接关系。然而，如果从该酶中除去ca2+，由于出现远程的结构变化，催化活性将丧失80%左右，但其剩余活性通常已足以降解在一般情况下污染酸制品的蛋白质。所以，蛋白酶k消 ...

问题可能原因解决方案回收率低膜结合液（MB）使用量不当按每1 mg凝胶或每1 µl DNA溶液加入1 µl膜结合液的比例（1:1）加入膜结合液溶胶不完全尽量将胶切成小块，加入膜结合溶液后于55～65℃加热助溶，确保溶胶彻底离心转速不够，溶液中的DNA没有与硅胶膜充分结合使用离心力≥12000x g的离心机，如达不到该转速，可通过适当延长离心时间确保胶溶液完全通过硅胶膜膜漂洗液（MW）未添加乙醇第一 ...

This protocol was designed to generate directionally end-labeled probes for DNaseI footprinting but it can be used for any application that requires end-labeled DNA probes.Solutions10 mM dNTP StocksTh ...

1. Add an equal volume (equal to sample volume) of P/C to sample.2. Mix (shake don't vortex).3. Take aqueous (upper) layer. (If dirty sample repeat Ph/Chl step until interface is fairly clean).4. Add ...