Nativechromatinimmunoprecipitationprotocol

实验概要The method is a native chromatin immunoprecipitation protocol.主要试剂1. 10 x TBS 0.1 M Tris-HCl (pH 7.5) 1.5 M NaCl 30 mM CaCl2 20 mM MgCl2 50 mM Na butyrate (pH 8.0)2. Digestion buffer 0.32 M sucrose 50 mM Tris-HCl (pH 7.5) 4 mM MgCl2 1 mM CaCl2 0.1 mM PMSF 5 mM Na butyrate3. Lysis buffer 1.0 mM Tris-HCl (pH7.4) 0.2 mM Na2EDTA 0.2 mM PMSF 5 mM Na butyrate4. Incubation buffer 50 mM NaCl 20 mM Tris-HCL (pH 7.5) 20 mM......阅读全文

Protocol-for-Trichl...

实验概要The  efficiency of nucleotide incorporation in DNA/RNA polymerization  reactions (e.g. transcription, reverse transcription, and DNA  replication)

Intracellular-Staining-Protocol

1. Fix cells- Add16% formaldehyde directly into culture medium to obtain a final concentration of 1.5% formaldehyde.2. Incubate in fixative for 10 min

Histone-blotting-protocol

实验概要 Western blot detection of histone proteins. 实验步骤 The  following protocol refers to the western blot detection of histone  proteins derived from p

Phycoerythrin-conjugation-protocol

Phycoerythrin conjugation protocolDavid's method modified from references (2) and (3). I used this method to conjugate a mouse IgG2a monoclonal an

Dot-blot-protocol

实验概要A  technique for detecting, analyzing, and identifying proteins, similar  to the western blot technique but differing in that protein samples are

Protocol-for-Cell-Fusion

Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, the

cDNA/AFLP-Protocol

Preparation of Para-magnetic beads from Promega cat#Z5482:a) suspend magnetic particles in bottle - transfer 200 ul (200 ug) of beads per sampleof RNA

Cell-Extraction-Protocol

实验概要Primary tissues  are valuable tools for the study of intracellular and extracellular  markers which characterize disease states. We have developed

Dot-Blot-Protocol

a. Label nitrocellulose membrane (using a pencil) to identify protein elution fractions.b . Pipette 2μl from each fraction onto the membrane, allow th

Basic-ELISA-Protocol

实验概要        There are many different types of ELISAs, which can detect the presence of protein in serum or supernatent. One of the most common typ

Nucleolar-Isolation-Protocol

We recommend that you first download and read this page as a PDF file. Using that as your guide, you can then follow the protocol below and view a Qui

BrdU-Labeling-Protocol

实验概要The thymidine analog, 5-bromo-2-deoxyuridine (BrdU),is a common reagent used for cell proliferation assays and for the detection of apoptotic

Cytotoxicity-Assays-Protocol

Cytotoxicity Assays ProtocolCell-mediated cytotoxicity was determined by using a standard microcytotoxicity assay. Briefly, target cells were pelleted

Sandwich-ELISA-Protocol

实验概要The  Sandwich ELISA measures the amount of antigen between two layers of  antibodies (i.e. capture and detection antibody). The antigen to be  mea

Western-Blotting-Protocol

实验概要The western blot  (sometimes called the protein immunoblot) is a widely used analytical  technique used to detect specific proteins in the given s

Tissue-Harvest-Protocol

TISSUES TO BE PROCURED(minimally and preferably within 5-8 hours after death):1. Brain2. Liver3. Muscle4. Skin5: Others as the specific case dictatesP

Western-Blot-Protocol

一、提取抗原蛋白将提取RNA途中留存的样品,加入150μl 100%酒精充分混匀,静置5min(RT), 2000×g , 4℃离心5min, 吸取上清至新管中, 加入750μl异丙醇, 混匀, 静置10min(RT), 12000×g, 4℃离心10min, 弃上清, 加入1ml 0.3mol/L

Colony-PCR-Protocol

1. Pull out eight glycerol stock plates from the –80oC freezer and set on bench top to thaw. Be sure to remove the foil seal before leaving the plates

Immunofluorescence-Microscopy-Protocol

实验概要Immunofluorescence  allows the imaging of a specific factor in cells or tissue sections  through the use of a specific antibody chemically which i

Adhesion-Assay-Protocol

Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Lamini

Protocol-for-dsRNA-Synthesis

实验概要        We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends

Protocol-of-Northern-blot

Protocol of Northern blot质粒的转化和扩增质粒的鉴定目的基因片段的切割3.1样品双酶切(175μl水解体系)DW 115μlBuffer B(10×) 17.5μlBaM H 15μlPst I 17μlDNA(MMP-9) 16μlBSA 4.5μl37℃水浴,3h。3.2

TAIL-PCR-Protocol

TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 react

RNA-Isolation-Protocol

RNA Isolation Protocol(Revised 5-15-2003)Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109 cells (OD600= 0.2 Dilute cells or scale up)

多名学者推荐全基因组分析转录因子功能新技术

  来自北卡罗来纳大学教堂山分校,NIH等处的研究人员利用一种新型方法,分析了酵母转录因子Rap1在整个基因组中的结合动态,从而可以更好研究这一转录因子的功能,这一方法将有助于科学家们实时分析转录情况,相关成果公布在Nature杂志上。   对于这一成果,来自法国国家科学研究中心的François

ELISA

Gangliosides ELISA protocol (Contributed by pingsunjim)This protocol can be used for detection of gangliosides.Specific antibodies to gangliosides are

染色质免疫沉淀法(Chromatin-immunoprecitation,ChIP)

染色质免疫沉淀法(Chromatin immunoprecitation,ChIP)是研究体内DNA与蛋白质相互作用的重要工具。它可以灵敏地检测目标蛋白与特异DNA片段的结合情况,还可以用来研究组蛋白与基因表达的关系。核小体组蛋白可以发生多种翻译后的共价修饰,如乙酰化、甲基化、磷酸化、泛素化等,这些

细胞组分和细胞器——染色体

Chromosomal DNA Prep : cultured cells/tissue samples (Mike A Dyer)This protocol was developed for cultured cells but should be appropriate for dissoci

新方法:超低细胞数表观基因组研究

  日本九州大学和东京理工学院的科学家们研发了一种使用非常少量的细胞,范围从100到1000的细胞,来分析DNA-蛋白质相互作用的新技术。他们的方法可以捕获前所未有的细胞内表观遗传组信息,该技术奖促进生物标记的发现并为精准医学开辟新道路。  这项被称为染色质整合标记测序(Chromatin Inte

Insect-DNA-Isolation-Protocol

实验概要The E.Z.N.A.® Insect DNA Kit is designed for efficient recovery of genomic DNA up to 60 kb in size from insects, arthropods, roundworms, flatw