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 transfection of suspension 293 human embryonic kidney cells (e.g. FreeStyle™ 293-F cells, Cat. no. R790-07) in defined, serum-free FreeStyle™ 293 Expression Medium, and is intended for use with the FreeStyle™ 293 Expression System (Cat. no. K9......阅读全文

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

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

Transfection-of-Mammalian-Cells-Using-Lipofectamine

Materials:        Lipofectamine Basal Medium containing 10% fetal bovine serum, 1% glutamine, 1% aa Basal Medium containing 1% glutamine

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

Transfection-of-siRNA-oligos-to-peritoneal-macrophages

Use DeliverX Transfection from PanomicsModified protocol belowStep 1:  Make complexesNote:  Do not scale up more than 3xThaw siRNA and DeliverX reagen

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

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

细胞转染(Cell-Transfection)技术综述

一、细胞转染途径转染大致可分为物理介导、化学介导和生物介导三类途径。电穿孔法、显微注射和基因枪属于物理介导技术;化学介导方法很多,如经典的磷酸钙共沉淀法、脂质体转染方法、和多种阳离子物质介导的技术;生物介导方法,有较为原始的原生质体转染,和现在比较多见的各种病毒介导的转染技术。1、物理介导(1)电穿

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

Cosmid-Cloning:-Cell-preparation,-DNA-packaging,-and-Cell-Transfection

Cosmid Cloning: Cell preparation, DNA packaging, and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction man

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

DNA转染

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

siRNA-转染程序

 A.siRNA转染的方法   哺乳动物转染的常见方法有:磷酸钙共沉淀、电穿孔法、DEAE-葡聚糖和polybrene、机械法(例如,显微注射和基因枪)、阳离子脂质体试剂,其中阳离子脂质体试剂转染法是目前最常用的转染方法。应用脂质体型转染试剂进行转染需要注重的几个方面:1.       转染试剂的用

表皮细胞的转染实验技巧

表皮细胞广泛遍布于身体,正常的表皮细胞较难转染,尤其是使用基于脂质体技术的转染试剂。我们使用电转(Amaxa)方法转染正常人的结肠表皮细胞并得到了65%的GFP标记细胞。非常感谢SignaGen,现在我们使用GenJet Ver II可以成功转染正常人的结肠表皮细胞并且转染效率显著提高至75%。

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

选择GenJetTM,LipoD293TM--PolyJetTMDNA转染试剂稀释溶液小技巧

选择何种稀释液稀释DNA及转染试剂,对于制备有效的转染复合物至关重要。除了温度,孵育时间外,稀释液的性质对于制备高效的转染复合物亦非常重要,同样影响DNA转染效率。根据我们的实验数据,使用合适的稀释液得到的转染效率是使用错误稀释液转染效率的至少50倍。更为重要的是,实验者总是忽视稀释液的重要性,甚至

如何优化GenMuteTM转染试剂,提高siRNA/DNA共转染效率?

GenMuteTM siRNA转染试剂(Cat#SL100568)是市场上最有力的siRNA传递工具之一。siRNA/DNA共转染时,siRNA的最佳浓度范围是0.5ηM到10ηM,过多的siRNA可能导致沉默效果差的“洪水效应”,切勿使用超过20ηM的siRNA。以24孔板为例,如下步骤将对如何优

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

siRNA体外转染——GenMuteTM-siRNA体外转染的优化

GenMuteTM siRNA转染试剂(Cat#SL100568)是市场上最有力的siRNA传递工具之一。最佳的siRNA浓度范围是1.0nM到10nM,过多的siRNA可能导致沉默效果差的“洪水效应”。我们实验室已经使用GenMuteTM转染试剂成功敲除了内源表达的生长因子。以24孔板为例,如下步

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

转染L929细胞的简单步骤

L929是小鼠成纤维细胞瘤细胞株,经常被用来检测TNF-alpha及TNF-beta。对TNF的处理经常会引发细胞凋亡及死亡,因此L929细胞经常被用作免疫分析。但是,转染L929细胞非常难,尤其使用基于脂质体技术的转染试剂,我们使用GenJet VerⅡ及PolyJet转染L929细胞获得了7

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

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

报告基因检测的精明之选—LipoD293DNA转染试剂

使用lipoD293转染试剂来转染哺乳动物细胞(比如Hela 、PC3),以期检测荧光酶报告基因,同其他转染试剂相比较,您会得到更多的荧光酶读数,更少的细胞死亡率。使用unsupplemented RPM11640/DMEM替代Opti-MEM培养基来稀释lipoD293及DNA(luciferas

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

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

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

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

xCELLigence系统实时监测低毒性XtremeGENE-DNA转染实验(二)

■  终点法分析,转染约72h后通过细胞增殖试剂盒WST-1分析确定细胞活性(图3)。与X-tremeGENE 9和X-tremeGENE HP Transfection Reagent相比,其他试剂转染细胞后,显著降低代谢活力,显示出细胞毒性效应。图3:比较罗氏X-tremeGENE 9和X-tr