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
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
Preparation－of－tubulin
Although many protocols for tubulin preparation are available, the procedure described below is the simplest and highest yielding preparation I have d
Template－Preparation
Template PreparationThe quality of sequencing results is directlyrelated to the quality of the template. ABI recommends a minialkaline-lysis/PEG preci
Platelet－Preparation
OUTLINEIn order to avoid platelet activation all manipulations must be performed as quickly and as acurate as possible.Work on ice if possible! This p
Liposome－Preparation
Liposome PreparationOBJECTIVE:Method for incorporating proteins into liposomes for liposome swelling assay or as an alternative antigen presentation m
SMEAR－PREPARATION
The preparation of a smear is required for many laboratory procedures, including the Gram-stain. The purpose of making a smear is to fix the bacteria
CAM－preparation
8 eggs per day, day 7- day 13 cut CAM, wash in precooled PBS,in 10 ml WASH 1 (PBS, 5 mM EDTA, COMPLETE) on icecut in pieces in petri dish on icecentri
Basic－procedures－for－bacteria－culture2
E. Elution of DNA fragments from agaroseDNA fragments are eluted from low-melting temperature agarose gels using an unpublished procedure first develo
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
酵母准备
Yeast DNA PreparationYeast Genomic Preparation (Gottschling Lab)Rapid method for yeast genomic DNA isolation Yeast DNA Preparation (rapid glass bead
PCR基本实验方法（五）
Cloning PCR ProductsT-A Cloning Strategy: Taq and other polymerases seem to have a terminal transferase activity which results in the non-templated ad
PCR基本实验方法（五）
Cloning PCR ProductsT-A Cloning Strategy: Taq and other polymerases seem to have a terminal transferase activity which results in the non-templated ad
Cloning－PCR－products－using－TA－vectors
Cloning PCR products using TA vectorsby Paul N. Hengen, Ph.D. *Methods and reagents is a unique monthly column that highlights current discussions in
Universal－RiboClone®－cDNA－Synthesis－System
Universal RiboClone® cDNA Synthesis SystemThe Universal RiboClone® cDNA Synthesis System contains the reagents required for the synthesis of double-st
热点课题研究之慢病毒载体的使用与技术展望（三）
1.ORF表达克隆产品 慢病毒货号 描述 滴度 体积及规格 慢病毒总量 LPP-货号-载体-100 ORF/Promoter/lncRNA 慢病毒 ORF： ≥ 10^8 T
细胞培养常规操作
常规操作(主要内容如下)· Aseptic Technique· Culture Vessels· Cell Counting· Primary Culture· Maintenance of Cell Line ·
荧光素酶检测（Luciferase－assay）
Introduction Luciferase can be used as a reporter gene to measure the activity of promoters, and/or the transfection efficiency. Aims You will be prov
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
FIXATION－and－DNA－Staining－for－Cell－Cycle－Analysis
BackgroundThis method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to ente
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
Southen杂交
Southern杂交One important thing for transfer:the weight of the object resting on top of the blotting apparatus should not exceed the weight equal to a 5
DNA电泳
DNA电泳（主要内容如下） Preparation of Agarose Gel and Electrophoresis Extraction of DNA From Agarose Gel Extraction of DNA from Acrylamide Gels DNA Marker
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
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
Rat－Liver－Preparation
实验概要The procedure presented below describes a method for preparing rat liver.主要试剂1. Aluminum Foil2. Liquid Nitrogen3. Dry Ice4. Ph
Metaphase－chromosome－preparation
Materials: RPMI 1640 medium fetal calf serum (FCS), 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best.-Nr. 295892) cell cuture flask
Preparation－of－human－platelets
Preparation of human platelets 1. Human blood was taken from drug-free volunteers on the day of the experiment using acidic citrate dextrose
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