CosmidCloning:Cellpreparation,DNApackaging,andCellTransfection
Cosmid Cloning: Cell preparation, DNA packaging, and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction manualHost bacteria (E. coli LE392)preparation:Pick one colony from fresh overnight culture on NZY agar plate and inoculate 50 ml NZY broth supplemented with 0.2% maltose and 10 mM MgSO4.Grow at 37C, shaking, 4-6 h (do not grow past OD600 1.0/ml).Pellet bacteria 2000 rpm f......阅读全文
Blood－Smear:－Preparation－and－Staining
Blood Smear: Preparation and StainingReference:Davidson, I. and Henry J., Clinical Diagnosis by Laboratory Methods, I. Davidsohn and J. Henry, eds., W
Preparation－and－Staining－of－Paraffin－Sections
I. Fixation and Processing of Tissue for Paraffin SectionsA. Fixation of Tissues in 10% Neutral Buffered FormalinSacrifice animal by prescribed and ap
巨噬细胞和单核白细胞
· Lymphocyte Transformation (Donis-Keller lab)Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocytes) from antico
细胞培养——细胞保藏
Working Cell Bank (Contributed by Nanci Donacki)Provides detailed protocol for establishing a working cell bank Master Cell Bank (Contributed by Na
Differential－cDNA－Screening－Procedures
Differential cDNA Screening ProceduresThe protocols listed refer to cDNA library construction and preliminary differential screening procedures. They
TOPO©－快速克隆技术介绍
Topo TA克隆方法（Topo TA Cloning Kit） Topo TA克隆原理与TA克隆一样，唯一不同的是TA克隆用的是T4连接酶把PCR片断连接到T载体上，而Topo TA Cloning用的是DNA Topoisomerase。 Topoisomerase的用途一般使用
RNA提取
RNA提取（主要内容如下）Tips for Handing RNA Total RNA IsolationmRNA IsolationrRNA IsolationOthersQ & A posted in the Method Forum Basic Procedures for Handing
ChIP－using－plant－samples
实验概要 The immunoprecipitation (IP) of cross-linked chromatin with antibodies specific for certain histone modifications (chromatin immunoprecipitati
Cell子刊：解析促癌“垃圾DNA”
来自德克萨斯大学健康科学中心癌症治疗与研究中心(CTRC)的研究人员，在乳腺癌形成的图画上又添加了一笔，在发表于8月12日《癌细胞》(Cancer cell)杂志上的一项新研究中，他们证实“远程雌激素反应元件”( distant estrogen response elements，DE
Cell：DNA修复必不可少的酶
最近，澳大利亚国立大学（ANU）和德国海德堡大学的科学家们，发现了DNA修复过程中的一个重要组成部分，将为开发新的抗癌药物打开大门。相关研究结果发表在10月27日的《Cell》杂志。本研究资深作者、ANU副教授Tamás Fischer说，这项研究发现，由DNA和RNA组成的混合结构，对于DNA损伤
Cell子刊：piRNA，DNA的贴身保镖
生殖细胞的DNA一直处于严密的保护之下，欧洲分子生物学实验室EMBL的研究团队日前为人们揭示了这个至关重要的保护机制，相关论文发表在七月九日的Cell Reports杂志上。 转座子又称为跳跃基因，是一种“自私”的DNA链。转座子能够进行自我复制，还能在染色体不同位点之间跳跃，会导致基因失活甚
Cell：DNA修复必不可少的酶
最近，澳大利亚国立大学（ANU）和德国海德堡大学的科学家们，发现了DNA修复过程中的一个重要组成部分，将为开发新的抗癌药物打开大门。相关研究结果发表在10月27日的《Cell》杂志。 本研究资深作者、ANU副教授Tamás Fischer说，这项研究发现，由DNA和RNA组成的混合结构，对于D
Genomic－Libraries
Genomic DNA libraries Size of some genomes and chromosomes:Comparative Sequence Sizes(Bases)(yeast chromosome 3)350 ThousandEscherichia coli (bacteriu
针对特定细胞选择更有效的实验步骤
GenJet Ver.II，LipoD293及PolyJet针对哺乳动物细胞的DNA体外转染试剂，均为您提供两种不同的转染步骤——一般步骤及高级步骤，这两种步骤分别针对不同的哺乳动物细胞。高级步骤主要针对难转的哺乳动物细胞，例如：MDCK，MDA-MB231，Caco-2等细胞。使用一般的转染步
Preparation－and－Staining－of－Frozen－Tissue－Sections
I. Preparation of Frozen Sections for SectioningMaterials neeed:2-methylbutane (isopentane)Liquid NitrogenDry icePeel-Away?/sup> base moldsFrozen tiss
Preparation－of－Broth－and－Plates,－etc.
Recipes: 1) LB BrothMake 16 gm of LB Broth Base (Gibco #M27800C) up to 800 ml in ddH2O. Swirl to dissolve, then add 110 µl of 10 N NaOH. Autoclave. 2
Large－Scale－Tubulin－Preparation——2
III. Pouring a 1L Phosphocellulose (PC) ColumnResin: Whatman P11 Cellulose Phosphate -- fibrous cation exchanger(1 gram of PC swells to about 4 ml pac
Preparation－of－Segmented－and－Polarity－Marked－Microtubules
Segmented and polarity-marked microtubules are very useful for many different types of in vitro assays. Segmented microtubules are microtubules with a
Preparation－of－Mitochondria－from－Rat－Liver
Preparation of Mitochondria from Rat LiverRat liver is an ideal source for functional intact mitochondria for a number of reasons. We use Sprague-Dawl
Preparation－of－Segmented－and－Polarity－Marked－Microtubules
Preparation of Segmented and Polarity Marked Microtubules Segmented and polarity-marked microtubules are very useful for many different types of in vi
Specimen－Preparation－for－Scanning－Electron－Microscopy
Specimen Preparation for Scanning Electron MicroscopyWe recommend consultation with one of the lab directors before preparing specimens. The methods p
Optimized－Method－for－the－Preparation－of－Rodent－Testicular－Cells2
Flow Cytometric AnalysisPrior to flow cytometric analysis, the vital dye Hoechst 33342 (Sigma-Aldrich, St. Louis, MO, USA) was added to cell suspensio
PCR产物克隆
PCR克隆主要有TA克隆法, 限制性酶切与连接法，杂交法和近期开发出来的特异性重组法等。但因为TA克隆法操作最简单，快速和高效的原因，成为Taq聚合酶PCR产物的最佳克隆方法。TA克隆法由Invitrogen发明，并拥有全球TA Cloning商标的ZL权。TA克隆方法（Original TA Cl
用CRISPR/Cas9对CART细胞进行多重基因编辑（三）
流式细胞术 Flow cytometry CytoFLEX (Beckman Coulter Inc) was used to perform fluorescent expression analysis. Cells were harvested on the following day
Pulse－Field－Electrophoresis
Manipulating and analyzing DNA are fundamentals in the field of molecular biology. Indeed, separating complex mixtures of DNA into different sized fra
PCR的下游应用
· Agarose Gel Electrophoresis of PCR Products (Robert H. Cruickshank)· Agarose Gel Electrophoresis of PCR Products (Immunology Resourc
Optimized－Method－for－the－Preparation－of－Rodent－Testicular－Cells1
Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the c
Coimmunoprecipitation－assays
co-IP assays can be performed between endogenous proteins or transiently or stably expressed exogenous - usually tagged - proteins. The advantage to u
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends2
3’ Adaptor Ligation and PurificationHeat shock the RNA by putting at 90°C for 30 seconds. Snap cool on ice.Set up the 3’Adaptor ligation reaction in a
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends3
Load your precipitated PCR samples into 2 consecutive lanes so as not to overload the lanes. For each different sample, I would run a separate ladder