TransformationofElectrocompetentE.coliwithBlue／Whiteselection
Desalt DNA template by EtOH precipitation in NaOAc followed by at least 2x washes with 70% EtOH. Resuspend in 5 - 15 µL of sterile H2O.Rinse cuvettes (if they have been used before) 5x with deionied H2O, and place them on ice. This is sufficient to avoid background growth in most cases.Set a BioRad MicroPulser to "Ec1" for 1 mm cuvettes, or "Ec 2" for 2 mm cuvettes.Electroporate the DN......阅读全文
ChIPChip－E.－coli
AbstractChIP-Chip stands for Chromatin Immunoprecipitation and chip in the sense of DNA microarray. It is a technique to determine the genome-wide bin
Sauer:Lysing－E.－coli－with－Lysozymes
Getting The Most Out Of Your BugsNative lysis is a staple protocol in practically every biochemistry lab, yet there is significant variability in the
Green－lab－protocol－for－vacuum－infiltration－transformation－of－Arabidopsis
This protocol is adapted from protocols by Nicole Bechtold (Bechtold et al. 1993), Andrew Bent (Bent et al. 1994) and Takashi Araki. No claims are
PEGMediated－Protoplast－Transformation－with－Naked－DNA
Direct introduction of DNA into plant protoplasts facilitates a rapid analysis of transient gene expression, as well as the generation of stably t
细菌转化（bacterial－transformation）原理和操作
1．目的学会质粒DNA转化感受态受体菌的技术。2．原理质粒DNA粘附在细菌细胞表面，经过42°C短时间的热击处理，促进吸收DNA。然后在非选择培养基中培养一代，待质粒上所带的抗菌素基因表达，就可以在含抗菌素的培养基中生长。3．器材旋涡混合器，微量移液取样器，移液器吸头，1.5ml 微量离心管，双面微
Nicotiana－(Nicotiana－tobaccum,－Nicotiana－benthamiana)
Agrobacterium -mediated transformation of Nicotiana species, namely, Nicotiana tobaccum and Nicotiana benthamiana , using leaf disks as the target e
Long－Term－Storage－of－Transformed－E.coli
Long Term Storage of Transformed E.coliTransfer 10 ml of 250 ml overnight culture in sterile flip-cap 15 ml Tube.Add sterile Glycerol to 15 % final co
Maxiprep－of－plasmid－DNA－from－E.－coli
IngredientsIngredients are per culture; make enough for one extra culture to allow for pipetting error).150μL sterile 50% glycerol1mL TEG (25mM Tris-C
Sauer:RNA－Purification－from－E.－coli
My Experience Purifying RNA from E. coliRegarding RNA extraction, there is a horrible tendency of people to use kits for RNA extraction with bacteria.
粘粒
Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures (Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to re
粘粒
Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures (Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to re
Bacterial－Colony－PCR
Bacterial Colony PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
细菌培养基
Preparation of LB Plate (Dr. Chastain)prepare LB plate with or without antibioticsBacterial Culture Media Recipes (WUGSC) M9 Plate Supplement (Gottsch
Construction－and－Manipulation－of－LargeInsert－Bacterial－Clone－Libraries
Acknowledgements The organizer of the workshop acknowledges Dr. Murray Milford, Professor and Interim Head, and Dr. Mark Hussey, Professor and Interim
Maxiprep－of－plasmid－DNA－from－E.coli－protocol
Solutions/reagents:LB broth + selective marker50% sterile glycerolTEG(25mM Tris-Cl, 10mM EDTA, 50mM dextrose)20 mg/ml lysozyme10% SDS4M NaOHautoclaved
Analysis－of－total－E.－coli－protein－by－SDS－PAGE
1. In microfuge tubes, spin down 0.1 ml of uninduced cells grown to near saturation or 0.15 ml of IPTG induced cells. Remove YT (or LB) media with a p
Targeted－Gene－Replacement－in－Fungal－Pathogens－via－Agrobacterium－...
Genome sequence data on fungal pathogens provide the opportunity to carry out a reverse genetics approach to uncover gene function. Efficient meth
重组DNA的分离、克隆与测序实验手册3
G. Bacterial cell maintenanceFour strains of E. coli are used in these studies: JM101 for M13 infection and isolation (4), XL1BMRF' (Stratagene) f
DNA转化实验指导4
2B. Transformation 1. Preparation of electrocompetent DH5a cells: autoclave 4 baffled 1 liter flasks containing 500 mL LB. Remove a 1 mL aliquo
重组DNA的分离、克隆与测序实验手册5
C. Random fragment end-repair, size selection, and phosphorylationSince both sonicated and nebulized DNA fragments usually contain single-stranded end
McKinney:TransformByElectroporation
Materials50mL 7H9 mycobacterial medium + 3mL per transformation102mL 10% glycerol (possibly a few mL more if you are doing many transformations)400mL
E.coli－Total－RNA－Labeling－Protocol－for－Spotted－Microarray
Note:Start with 20 ug of total RNA for each labeling reaction.All solutions that can be filtered should be filtered.Cy dyes are light sensitive and sh
Genetic－Transformation－of－Cotton－with－a－HarpinEncoding－Gene－hpaXoo－Conf...
Genetic Transformation of Cotton with a Harpin-Encoding Gene hpaXoo Confers an Enhanced Defense Response Against Verticillium dahliae KlebThe soil
酵母转化的几种方法
Modified Yeast Transformation Inoculate cells from an overnight culture into 50 ml YEPD and incubate at 30°C with shaking. Typically, add 0.1 to 0.2 m
超级感受态细胞的制备
The Inoue Method for Preparation and Transformation of Competent E. coli: "Ultra Competent" CellsJoseph SambrookPeter Maccallum Cancer Institute and T
DNA转化实验指导1
CONTENT Transformation-Competent E. coli preparation Inoue "ultra-competent" methodRubidium chloride methodCosmid packaging protocol DNA Ligation an
技术和方案26－小量提取E.coli－DNA
试剂、试剂盒Terrific broth实验步骤1.将单菌落接种于 3 ml 含有 100 ug/ml 氨节青霉素的 Terrific broth 培养基中，37°C 培养过夜。2.取 1.5 ml 细胞悬液于微量离心管中，13000 r/min 离心 1min，弃去上清。3.加入 100 ul 溶
E.coli基因文库的分类和选择
自 1995 年成立以来，Dharmacon 在生物信息学，RNA 生物学和合成化学方面的专长 , 使我们能够开发出一整套研究基因功能的产品。作为 RNA 定制合成的leader，Dharmacon 公司是 RNA 干扰新发现领域的早期参与者，并且在若干重要的科学发现中，以及确保沉默效率的
E.coli基因文库的分类和选择
文库种类Dharmacon 大肠杆菌资源库包括大肠杆菌Keio基因敲除文库，监测基因表达的启动子-GFP融合文库，以及用于研究蛋白质-蛋白质相互作用的标记ORF文库。大肠杆菌（E.coli）基因cDNA&ORF文库 （1）大肠杆菌Keio基因敲除文库（E.coli Keio Knockout Col
基于epMotion－5075t系统与KPPA－HyperPlus试剂盒的全自动测序..1
基于epMotion 5075t系统与KPPA HyperPlus试剂盒的全自动测序前文库制备方案Automated KAPA HyperPlus DNA Library Preparation for Illumina® Sequencing on the Eppendorf epMotion®