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
Method:－Preparation－of－Lymphocyte－Cell－Pellet－for－Storage
Method: Preparation of Lymphocyte Cell Pellet for Storage June 10, 1990 Rosalie Veile Purpose: Following propagation to 1 X 108 cells, l
Human－Peripheral－Blood－Mononuclear－Cell－Preparation
This protocol describes a procedure for isolating human peripheral blood mononuclear cells (lymphocytes and monocytes) from a Buffy Coat (obtained fr
Preparation－of－Agarose－Gels－for－DNA－separations
Weigh out the desired amount of agarose and place in an Erlenmeyer flask with a measured amount of electrophoresis buffer, e.g. for an 0.8% gel, add 0
DNA体外转染试剂_如何准备转染所需的质粒DNA
转染效率受到诸多因素的影响，除了细胞、培养基和载体等影响因素外，另外一项非常重要的因素便是DNA的质量。为了比较不同厂家的转染效率，我们分别从三家不同的供应商购买了质粒制备试剂盒来制备pEGFP-N3质粒DNA，并通过不同的方法将制备的pEGFP-N3质粒转运至NIH-3T3细胞。pEGFP-N3质
Large－Scale－Plasmid,－Cosmid,－BAC,－PAC,－and－Fosmid－DNA－Isolation
DNA Isolation by a Cleared Lysate Method Followed by Double Acetate Precipitation Version 3b - updated September 26, 1999The Most Recent Roe Lab Imple
胚胎干细胞培养技术大全
MEDIA AND SOLUTIONS REQUIRED FOR ROUTINE ES CELL CULTURE Routine Culturing of ES Cells ISOLATION OF PRIMARY MOUSE EMBRYO FIBROBLASTS MITOM
Preparation－of－cytospin－from－single－cell－suspension.
Cell number, speed and trivial details affect cytospin . Basic protocol: Prepare a cell suspension of not more than 0.5 x 106 cells / ml of protein
噬菌体的生长
Preparing Lawn Cells for M13 Cloning (Life Technologies)Lawn cells require the F' episome for M13 infection and may be prepared Streaking Lambda
Preparation－of－Yeast－DNA－Embedded－in－Agarose－Plugs
Preparation of Yeast DNA Embedded in Agarose PlugsAnja van Brabant(adapted from Iadonato, S. P., and A. Gnirke. 1996. RARE-cleavage analysis of YACs.
Column－Method－for－Lambda－Phage－DNA－Preparation
Purpose:Mini-prep method for lambda phage DNA purification from lysates.Time required:4 hours once the lysate is in handSpecial supplies required:BioR
Cosmid提取技术
Day OneStart overnight cultures of Cosmid in fresh L-Broth + 25 ug/ml kanamycin. 4 x 250 ml each, inoculate each with metal loop from frozen glycerol
Transfection－with－PEI
实验概要Use PEI (Linear 25 kDa Reagent) to transfect in HEK293T cells.主要试剂PEI is polyethyleimine, a 25 kDa linear from Polysciences Inc. Make up solutio
细胞遗传学——原位杂交(ISH)
In Situ Hybridization· In Situ Hybridization (jsmith1@po-box.mcgill.ca)In situ hybridization, as the name suggests, is a method of localizing,
质粒的大量制备
· Plasmid Mini and Maxi Prep Methods (Gimila Lab) · Maxi-preps and all media, solutions (NWFSC)Isolation of cosmid, plasmid and P1 DNA
质粒的大量制备
· Plasmid Mini and Maxi Prep Methods (Gimila Lab) · Maxi-preps and all media, solutions (NWFSC)Isolation of cosmid, plasmid and P1 DNA
Method:－Preparation－of－Lymphoblastoid－Cell－Lines－for－Long－Term－Storage
Method: Preparation of Lymphoblastoid Cell Lines for Long Term Storage May 30, 1990 Rosalie Veile Purpose: To store cell lines in a form
Hematopoietic－Stem－Cell－Targeting－with－Surface1
Hematopoietic Stem Cell Targeting with Surface-Engineered Lentiviral VectorsEls Verhoeyen and François-Loïc CossetAdapted from Gene Transfer: Delivery
Fusion－and－Cloning
Author: Nanci DonackiSource: Contributed by Nanci DonackiAbstract: Procedure for establishing hybridoma in one stepReagents(StemCell Technologies, Inc
Fusion－and－Cloning
ReagentsMedium A - Pre-fusion Medium and Hybridoma Expansion MediumMedium B - Fusion Medium Medium C - Hybridoma Recovery MediumMedium D - Hybridoma S
High－Molecular－Weight－Yeast－Liquid－DNA－Preparation
Purpose: To isolate intact, high molecular weight DNA from yeast cells for subcloning and rare cutting restriction enzyme analysis. One can expect
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Minipreparation
实验概要Plasmid DNA is isolated from small-scale (1-2 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and Solutions: Alkaline lysis
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Maxipreparation
实验概要Plasmid DNA is isolated from large-scale (500 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and SolutionsAlkaline lysis solu
细菌人工染色体
The Construction of Bacterial Artificial Chromosome (BAC) Libraries (complete manuscript) (Clemson University Genomics Institute) Construction of BAC
A－practical－method－for－the－preparation－of－total－DNA－from－filamentous－fungi
Most methods of DNA preparation from fungi are time-consuming due to the need to first make protoplasts, expensive for chemicals such as cesium chlori
General－Cloning－Protocols
Large Scale Preps: (See Large scale plsasmid prep protocol for more details) Cultures: Inoculate a 5 mL LB/Amp (50 - 100 µg/mL) culture in early a
DNA的酶学操作
DNA的酶学操作DNA Modifying Enzymes (Michael Blaber)Introduction to bacterial restriction/modification system. It provides very useful background knowledge
Streptomyces:Protocols/Conjugation
Intergeneric Conjugation and Overlay Description Transfer of plasmid/cosmid DNA from a host strain, e.g. E.coli ET12567 [pUZ8002], to the recipient
Preparing－Antibiotics－Stock－Solution－and－Ampicillin－Agar－Plates
AMPICILLIN Beta-lactam-antibiotics are not very stable when dissolved. Slow but steady degradation happens even when frozen to -20°C. Therefore comme
293fectin™－Transfection
实验概要293fectin™ is a proprietary, cationic lipid-based formulation for transfection of DNA into eukaryotic cells. 293fectin™ is optimized for transfe
哺乳动物RNAi技术－Mammalian－RNA－Interference
Mammalian RNA InterferenceThomas TuschlLaboratory for RNA Molecular BiologyThe Rockefeller University, New York Excerpted from RNAi: A Guide To Gene S