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
Preparation－of－Mouse－Neutrophils
实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline solution [HBSS; Invitrogen, Grand Isla
HELPER－PHAGE－PREPARATION
HELPER PHAGE PREPARATION1. Grow an overnight of NM522 in NZCYM medium.2. Dilute overnight 1:100 and grow to an A600 = 0.3 (@2.5 x 108 cells/ml).3. Inf
Preparation－of－Mouse－Neutrophils
实验概要Preparation of Mouse Neutrophils实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline so
DGK－Membrane－Preparation
Reagents:Bacterial strainE. coli N4830/pJW10LB amp media50 µg/ml ampicillinHigh salt bufferfor 1 L50 mM KH2PO4 6.8 g150 mM KCl 11.18 g50 mM sodium pyr
PREPARATION－OF－SEQUENCING－GELS
MATERIALS:2-glass plates1 sharks -tooth comb and spacersWhatman 3 mm paper30 or 40% acrylamide-bis (19:1)10X TBEurea10% ammonium persulfateTEMED60 cc.
转基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Core)Thi
如何优化PolyJet转染试剂？
我们使用PolyJet DNA转染试剂转染表皮细胞及Raw267.4细胞非常有效，并且毒性很小。对于如何更好的优化PolyJet转染试剂，我乐意分享以下几点：1、DNA的质与量。DNA的纯度对于转染实验至关重要。由E Coli制备的DNA，A260/280必须达到1.80甚至更高。对于6孔板，通常每
A－rapid,－quantitative－and－inexpensive－method－for－detecting－apoptosis2
Figure 1.Determination of apoptosis in transiently transfected murine [beta] tumor cells. (A) The number of apoptotic [beta]HC 13T tumor cells (% apop
BLOCKiT™－Fluorescent－Oligo－as－RNAi－Transfection－Control
实验概要Intended UseDynabeads® Streptavidin are ideal for numerous applications, including purification of proteins, nucleic acids purification, protein
The－Dos－and－Donts－of－Cell－Culture
Given below are a few of the essential "do’s and don'ts" of cell culture. Some of these are mandatory e.g. use of personal protective equipment (P
Sequencing－off－Cosmid,－BAC,－PAC,－－with－ABI－Big－Dye－Terminators
Big Dye Protocols and Notes - Cosmid, BAC, BAC, Fosmid TemplatesHi all,Over the past two months, we have been testing various reaction conditions for
siRNA用户手册
The siRNA user guide (revised May 6, 2004) Selection of siRNA duplexes from the target mRNA sequenceUsing Drosophila melanogaster lysates (Tuschl et
SiRNA用户指南
Selection of siRNA duplexes from the target mRNA sequenceUsing Drosophila melanogaster lysates (Tuschl et al. 1999), we have systematically analyzed t
96well－Plate－Dual－Luciferase－Assay96孔板荧光素酶检测
Reagents: Plasmids: can be found in my plasmid(1) box in the ?20oC freezer. 985 : FR-tk-luciferase 2517 : Renilla-tk-luciferase 2145 : GFP Lipfectami
ChIP－using－plant－samples－–－Arabidopsis
实验概要This protocol describes how chromatin is prepared from Arabidopsis, which can subsequently be used for chromatin immunoprecipitation (ChIP). T
Cell：超越DNA的遗传与编程
来自犹他大学Huntsman癌症研究所(HCI)的研究人员在新研究中发现，父源基因在受精之时已预编程至胚胎所需状态，而母源基因则处于另一种状态，还必须进行重编程才能与之相匹配。这一研究发现对于发育生物学和癌症生物学均具有极其重要的意义。研究论文发表在5月9日的《细胞》(Cell)杂志上。
Cell：DNA损伤应答的新通路
加州大学的科学家们在研究DNA损伤对高尔基体的影响时，发现了DNA损伤激活的新通路，这一通路影响着机体中细胞对化疗的应答。 包括化疗和放疗在内的标准癌症治疗策略，通过诱导细胞出现DNA损伤起作用。DNA损伤启动的信号通路会导致细胞死亡，人们正是在这一机制的基础上消灭癌细胞。更好地理解这些细
Cell：DNA损伤揭示抗癌新疗法
大自然中每一个有机体都会不惜代价保护自身的DNA，但细胞如何精确区分自身DNA的损伤还是入侵病毒外源DNA的损伤依然是个谜底，近日刊登在国际杂志Cell上的一篇研究论文中，来自索尔克研究所的研究人员通过研究揭示了细胞反应系统精确区分上述两种威胁的机制，相关研究或可帮助开发新型的癌症选择性病毒疗法
《Cell》探讨“垃圾”DNA的新功能
美国国立卫生研究院下属国家关节肌肉骨骼及皮肤病研究所(NIAMS)的科学家们在新研究中证实，过去被认为是“垃圾”的DNA在免疫系统反应中发挥了关键性的作用。研究结果发表在近期的《细胞》(Cell)杂志上，有可能促成发现治疗免疫相关疾病的新靶点。 人类基因组中有32亿DNA碱基对，但只有2%
Cell：“暗物质”DNA影响大脑发育
十多年来，由基因组中的“暗物质”片段（没有明显功能的缠绕在一起的DNA长链）带来的谜题一直困扰着科学家。如今，一个团队最终破解了这个谜题。 这个谜题集中在不编码蛋白质但在很多动物中保持相同的DNA序列。通过删除其中一些“超保守元素”，研究人员发现，这些序列能微调编码蛋白质的基因表达，进而指导大
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
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
Sample－preparation－(analytical－gels)
Sample preparation and solubilization are crucial factors for the overall performance of the 2-D PAGE technique. Protein complexes and aggregates shou
Preparation－of－Lactobacillus－Competent－Cells
OverviewInstructions on how to prepare Lactobacillus plantarum competent cells before electrotransformation.MaterialsMRS mediaCulture of L. plantarum
Large－Scale－Tubulin－Preparation
Tubulin is purified from bovine/porcine brain by two cycles of polymerization/depolymerization followed by removal of copurifying proteins on a phosph
Easy－YAC－Preparation－Method
YAC TRANSFORMATION OF C. ELEGANS USING TOTAL YEAST GENOMIC DNA[This method is described in The Worm Breeder's Gazette (1997) A. Davies and J. Shaw
Preparation－of－nucleic－acid－probes
Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as s
Protocols－for－LCM－preparation－and－analysis
Protocols for LCM preparation and analysis I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue SectionsA. EmbeddingB. CuttingC. StainingII. Pr
Preparation－of－tubulin2
DAY 2: Cycling preparation of MT protein.Keep the brains in an evacuated plastic ziplock bag buried in ice from the time of slaughter during transport
Tissue－preparation－protocol－for－ChIP
实验概要This protocol describes how chromatin is prepared from tissue, which can subsequently be used for chromatin immunoprecipitation (ChIP). It is re