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
浙江大学Cell子刊解析DNA损伤修复
维持基因组DNA的稳定对于细胞存活和肿瘤抑制至关重要。浙江大学生命科学研究院的科学家们对DNA修复机制中的一个重要复合物进行了研究，阐述了该复合体组装和发挥作用的结构基础。文章于三月十三日发表在Cell旗下的Cell Reports杂志上。 细胞基因组的完整性，持续受到多种内外因素的挑
《Cell》子刊：如何减缓肿瘤进展？利用DNA修复！
DNA修复机制在肿瘤的发生和进展过程中其到关键的作用。因此，研究人员不断探索靶向DNA修复机制关键蛋白在肿瘤疗中的作用。目前研究人员提出了对抗备用DNA修复机制的新方法。 DNA修复关键工匠-BRCA蛋白 被称为BRCA的蛋白质是BReast CAncer易感基因的缩写 ，这种蛋白在细胞DN
Cell：打破常规！发现独特的DNA编辑功能
一种单细胞纤毛虫物种以一种看似不可能的方式使用细胞转录复合体。在一项新的研究中，来自瑞士伯尔尼大学的研究人员首次详细地描述了 “垃圾DNA”在遭受降解之前转录的机制。这种机制是非常巧妙的。 它听起来像是一场设计竞赛中的获奖设计方案：当小片段信息太短而不适合放入读取设备中时，如何读取这些小片段信
Cell：DNA断裂与大脑学习及老化相关
来自MIT的研究者们发现，神经元活性可以快速大量激活一些早期反应基因，从而促进大脑的学习和记忆。DNA双链断裂的形成，是一种生理活动，可以快速解除拓扑蛋白对早期反应基因的抑制，从而在神经元中表达这些早期反应基因。这项研究结果，有助于研究人员们研发出新的方法，防治认知下降疾病，例如老年痴呆症。此项
Cancer－Cell：隐藏在“垃圾DNA”中的癌基因
科学家们一直致力于在基因组中寻找与癌症有关的遗传学改变。在此基础上，人们对癌症的理解以及癌症治疗都取得了巨大的进展。 这类研究大多关注基因组的编码区域，但这一区域只占到人类DNA的2%，应该还有大量癌症相关突变发生在非编码DNA上。非编码DNA曾被称为“垃圾DNA”，虽然人们早已知道它们其实并
Genomic－DNA－Extraction－－PureLink™
实验概要The PureLink™ Genomic DNA Purification Kit allows rapid and efficient purification of genomic DNA. The kit is designed to efficiently isolate g
Measurement－of－Green－Fluorescent－Protein－Expression
ReagentsCells to be studied expressing green fluorescent protein (GFP). Note that the same cell type without GFP is needed as control.Hoechst 33342 s
The－informationprocessing－pathway－at－the－IFNbeta－enhancer
The packaging of eukaryotic DNA into nucleosomes inhibits the access of factors to DNA and results in the repression of transcription, replication and
胚胎干细胞培养技术大全
MEDIA AND SOLUTIONS REQUIRED FOR ROUTINE ES CELL CULTURERoutine Culturing of ES CellsISOLATION OF PRIMARY MOUSE EMBRYO FIBROBLASTSMITOMYCIN C TREATMEN
Creation－and－Use－of－Infectious－Virus－Vector
Creation and use of your infectious vector:Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 ml of media. (This can be scaled up if desired).The f
CREATION－AND－USE－OF－YOUR－INFECTIOUS－VECTOR
实验概要CREATION AND USE OF YOUR INFECTIOUS VECTOR实验步骤Day 1 1. Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 mL of media. (This can be scal
流式细胞仪技术专辑
Flow Cytometry Analysis (Springer Lab, Harvard University) Flow cytometry employs instrumentation that scans single cells flowing past excitation sour
pEGFPC1质粒图谱及信息
pEGFP-C1Restriction Map and Multiple Cloning Site of pEGFP-C1. (Unique restriction sites are in bold.) The Xba I and Bcl I sites (*) are methylated in
pEGFPN1质粒图谱及信息
多克隆位点区Restriction Map and Multiple Cloning Site of pEGFP-N1. (Unique restriction sites are in color or bold.) The Not I site follows the EGFP stop cod
pEGFPC2质粒图谱及信息
多克隆位点区（MCS）Restriction Map and Multiple Cloning Site of pEGFP-C2. (Unique restriction sites are in color or bold.) Note that the Eag I site is not uni
组织学——组织制备
· Histological techniques (William H. Heidcamp)Very detailed guide to histological techniques, like fixation, dehydration, embedment and subs
Chromatin－Remodeling－by－hSWI/SNF－ATPdependent－Complexes
The eukaryotic genome is packaged by histone and nonhistone proteins to form chromatin. The assembly of nucleosomesas well as compaction of nucleosoma
Western－杂交
Western 杂交（主要内容如下）Preparing of Protein LysatesWestern BlottingFar Western BlottingSemi Dry BlottingStripping MembranesTrouble Shooting and OthersPrepa
PREPARATION－OF－2％－FORMALDEHYDE－STOCK－SOLUTION－(2－METHODS)
METHOD 1:Formaldehyde preservative – 2% formaldehyde solution in protein-free phosphate-buffered saline (PBS).Prepare as follows:Add 2 g paraformaldeh
96Well－Sample－Preparation－for－Suspension－Cells
实验概要The procedure presented below describes a facile method for studying signal transduction events with suspension cells (Jurkat, Raji, THP-1, etc.
96Well－Sample－Preparation－for－Suspension－Cells
实验概要The procedure presented below describes a facile method for studying signal transduction events with suspension cells (Jurkat, Raji, THP-1, etc.
96Well－Sample－Preparation－for－Adherent－Cells
实验概要The procedure presented below describes a facile method for studying signal transduction events with adherent cells (HeLa, MCF-7, BALB/c 3T3, et
GOLGIVESICLE－PREPARATION－FROM－PEA－HYPOCOTYLS
PREPARE SOLUTIONS1. 0.25 M Sucrose Solution:Mix 40 g of sucrose (0.25M), 50 mL of 1M KH2PO4, pH 6.65 (0.1M), 2.5 mL of 1M MgCl2 (5 mM), and dH2O to 50
Preparation－of－Stroma,－Thylakoid－Membrane,－and－Lumen－Fractions－from－...
Preparation of Stroma, Thylakoid Membrane, and Lumen Fractions from Arabidopsis thaliana Chloroplasts for Proteomic AnalysisFor many studies regarding
Preparation－of－denaturing－6％polyacrylamide－gels－for－microsatellite－analysis
Preparation of denaturing 6% polyacrylamide gels for microsatellite analysis(also for SSAP, high-resolution IRAP/ISSR and other analyses)Saadiah Jamli
Preparation－of－Low－Density,－CollagenaseDigested－Splenocytes
1. Dilute 2 ml of 4000 U/ml Collagenase D as follows: 1 ml into 9 ml HBSS/Ca++/Mg++ (=400U/ml) and 1 ml into 39 ml HBSS/Ca++/Mg++ (=100U/ml). Put on i
96Well－Sample－Preparation－for－Adherent－Cells
实验概要The procedure presented below describes a facile method for studying signal transduction events with adherent cells (HeLa, MCF-7, BALB/c 3T3, et
96Well－Sample－Preparation－for－Suspension－Cells
实验概要The procedure presented below describes a facile method for studying signal transduction events with suspension cells (Jurkat, Raji, THP-1, etc.
ASENSITIVE－METHOD－FOR－DETECTION－OF－APOPTOSIS－BY－SINGLE－LASER－FLOW－CYTOMETRY
MATERIALS:1. 1 X PBS (PBSAz, 1 X PBS, e.g., Irvine Scientific, CA, containing 2% newborn calf serum and 0.1% sodium azide)2. 7-Amino-actinomycin D (7-
DNA－methyltransferase－Assay
Methylated CpG island Amplification Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polymerase