TransfectionofMammalianCellsUsingLipofectamine

Materials: LipofectamineBasal Medium containing 10% fetal bovine serum, 1% glutamine, 1% aaBasal Medium containing 1% glutamineBasal Medium containing 20% fetal bovine serum, 1% glutamine, 1% aa1. In a six-well or 35 mm tissue culture plate, seed ~2x 105 cells per well in 2 ml Basal Medium containing 10% FBS.2. Incubate the cells at 37C in a CO2 incubator until the c......阅读全文

Transfection-of-Mammalian-Cells-Using-Lipofectamine

Materials:       LipofectamineBasal Medium containing 10% fetal bovine serum, 1% glutamine, 1% aaBasal Medium containing 1% glutamineBasal Medium cont

DNA转染

DNA转染·         Transfection of Mammalian Cells Using Lipofectamine (LTI)·         Guide to Eukaryotic Transfections with Cationic Lipid Reagents (PDF)

Transfecting-Plasmid-DNA-into-NIH3T3-Cells-Using-Lipofectamine™-LTX-Reagent

实验概要Lipofectamine™  LTX Reagent is a proprietary, animal-origin free formulation for the  transfection of DNA into eukaryotic cells with low cytotoxic

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

Transient-transfection-into-293T-cells

PurposeTransient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) pro

Transient-Transfection-of-Cos1-Cells

Transient Transfection of Cos-1 CellsNote: This protocol has been optimized for Cos-1 cells. Successful transfection of each cell type requires optimi

Enzyme-Kinetics-assay-of-the-WT

To assay 17 b-HSD activity in lysates, cells were harvested 48h after transfection using PBS Enzyme Free Cell Dissociation Solution ( specialty Media

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

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

FACS-Analysis-Using-Peripheral-Blood-Cells

FACS Analysis Using Peripheral Blood CellsCollect blood (75 microliters) into 1ml PBS containing 5 microM EDTA (10 microliters of 0.5 M stock) and mix

Detection-of-MicroRNA-Heterogeneity-in-Single-Cells-Using-an-Automated

Introduction  MicroRNA  (miRNAs) are short (18–24 nucleotides), non-coding RNAs that regulate  gene expression by both disrupting messenger RNA (mRNA

Growth,-Maintenance-and-Transfection-of-Suspension-Adapted-293EBNA-cells

ProcedureI. INTRODUCTIONThe 293 EBNA cell line is established from primary embryonal human kidney cells transformed with sheared human adenovirus type

用CRISPR/Cas9对CART细胞进行多重基因编辑(二)

细胞系 Cell linesThe following CD19-expressing immortalized cell lines were used: Raji (Burkitt’s lymphoma cell line, ATCC-CCL86),Daudi (B lymphoblast ce

Identification-of-a-Mutant-Kinase/ATP-Analog-Pair2

Cellular Transfection and Immunoprecipitation  Before proceeding with the experiments outlined below, all kinase pocket mutants should be characterize

Reverse-Transfection-for-Gene-Function-Analysis

This guide describes a microarray-based system for the functional analysis in mammalian cells of many genes in parallel. Mammalian cells are cultured

如何优化LipoD293转染试剂?

LipoD293DNA转染试剂是脂质体DNA转运工具的升级版本。我们实验室使用LipoD293DNA转染试剂成功转染了HepG2,LNCaP,CHO及HEK293细胞。接下来,我们乐意就如何提高转染效率,降低毒性等方面的小技巧同大家分享。1、LipoD293/DNA的比率。尽管合适的比率由细胞类型决

通过细胞受体代谢生物素化进行图像分析

Metabolic biotinylation of mammalian cell receptors for imagingBakhos A. Tannous , btannous@hms.harvard.edu, Massachusetts General Hospital and Harvar

Hematopoietic-Stem-Cell-Targeting-with-Surface1

Hematopoietic Stem Cell Targeting with Surface-Engineered Lentiviral VectorsEls Verhoeyen and François-Loïc CossetAdapted from Gene Transfer: Delivery

NAi-protocol

siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western

RNAi-protocol

 siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western

siRNAs结合生物芯片的实验设计2

Figure 2. Silencer ™ siRNA Validation Data Generated Using Applied Biosystems TaqMan® Gene Expression Assays. The indicated Silencer Validated siRNAs

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

Green-Fluorescent-Protein-as-an-Indicator-ofTransfection-in-Chicken-Embryos

Green fluorescent protein (GFP) is responsible for the bioluminescence of the Pacific Northwest jellyfish, Aequorea victoria. In A.victoria, the 27-kD

Transfecting-Suspension-Cells

实验概要将转移基因整合到细胞染色体DNA上,形成稳定表达转移基因的细胞系。 实验原理    细胞转染技术是目前广泛应用于病毒基因结构与功能以及基因调控等的研究。细胞转染可分为短暂转染和稳定(或永久) 转染两种。在短暂转染中,被转染基因并不整合至细胞染色体中,因而不能随细胞分裂而传代。转入病毒基因的转

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

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

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

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