DAPICounterstainingProtocols
實驗概要The blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it appears to associate with AT clusters in the minor groove. Binding of DAPI to dsDNA produces a ~20-fold fluorescence enhancement, apparently due to the displacement of water molecules from both DAPI and the minor groove. DAPI also binds RNA, however in a different binding mode—one thought to involve AU......閱讀全文
DAPI－Counterstaining－Protocols
實驗概要The ?blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; ?it appears to associate with AT clusters in the minor groove. Binding
DAPI－Nucleic－Acid－Stain
實驗概要The ?blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it ?appears to associate with AT clusters in the minor groove. Binding
LCM－PROTOCOLS
Slide SectioningParaffin blocks-?For?DNA?analysis:Special LCM processing schedule is followed.The water bath is cleaned using RNAse Zap?, rinsed thoro
Neutralizing－Bioassay－Protocols
Neutralizing Bioassay ProtocolsIntroductionAntibodies that block binding of cytokines to their specific receptors and neutralize their effects are cri
Streptomyces:Protocols/Conjugation
Intergeneric Conjugation and OverlayDescription?Transfer of plasmid/cosmid DNA from a host strain, e.g. E.coli ET12567 [pUZ8002], to the recipient str
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.m. w
CGH－Protocols－（四）
CGH Image acquisitionImages were acquired through a Zeiss Axiophot fluorescence microscope using a Plan NEOFLUAR oil objective x63, N.A. 1.25 (Zeiss,
Streptomyces:Protocols/PCR
Description?Polymerase Chain Reaction (PCR) is a method of amplifying a specific DNA target sequence. The cycle involves denaturing the template doubl
CGH－Protocols－（二）
DNA preparation by cryotom tissue dissectionPreparations/Materials:?Cool cryostat down to -20 to -30°C about 3 hours prior to dissection?Label eppendo
CGH－Protocols－（一）
Metaphase chromosome preparationMaterials:?RPMI 1640 medium?fetal calf serum (FCS), 20%?Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best
Smolke:Protocols/Western
OverviewBlotting for large V5-tagged proteins in?S. cerevisiaeMaterialsY-PER (Pierce)Halt EDTA-free Protease Inhibitor (Pierce)NuPAGE Novex Bis-Tris 4
Western－Blotting－Protocols
back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.
CGH－Protocols－（三）
Hybridizationreagents:?labeled tumor and normal-DNA (see protocol Nick translation)?salmon sperm DNA, 10 mg/ml (e.g. Promega)?human Cot1 DNA, 1 mg/ml
DAPI特點介紹
DAPI 是一種能夠與DNA強力結合的熒光染料。它結合到雙鏈DNA小溝的AT堿基對處，一個DAPI分子可以占據三個堿基對的位置。結合到雙鏈DNA上DAPI分子的熒光強度提高大約20倍，常用與熒光顯微鏡觀測，根據熒光的強度可以確定DNA的量。另外，因為DAPI可以透過完整的細胞膜，它可以用于活細胞和固
什么是DAPI？
DAPI，即4',6-二脒基-2-苯基吲哚，是一種能夠與DNA強力結合的熒光染料，常用于熒光顯微鏡觀測。因為DAPI可以透過完整的細胞膜，它可以用于活細胞和固定細胞的染色。
DAPI染色流程
DAPI染色1．原理DAPI為4’，6二脒基-2-苯吲哚(4’，6―diamidino-2―phenylindole)能與雙鏈DNA小槽，特別是AT堿基結合，也可插入少于3個連續AT堿基對的DNA序列中。當它與雙鏈DNA結合時，熒光強度增強20倍，而與單鏈DNA結合則無熒光增強現象，因此是一種簡易、
Protocols－for－LCM－preparation－and－analysis
Protocols for LCM preparation and analysis?I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue SectionsA.?EmbeddingB.?CuttingC.?StainingII. Pr
Streptomyces:Protocols/Spore－Prep
Spore Prep - Inoculating & HarvestingDescription?A spore prep is a method of preserving a sporulating strain of Streptomyces. The stock is stored in 2
ORNL－MICROARRAY－HYBRIDIZATION－PROTOCOLS
Direct labeling of total RNA with Cy3 and Cy5:A. MATERIALSRNeasy? Mini Kit (Qiagen; Cat # 74106)?SuperScript II RT (200U/μL) (Life Technologies; Cat #
Rat－Blood－Collection－Protocols
實驗概要The procedure presented below describes a method for collecting rat blood.實驗步驟Rat should be fully anesthetized (e.g., unresponsive to toe pinch).1
Streptomyces:Protocols/Transformation－by－Electroporation
Description?Transform?E.coli?cells with plasmid/cosmid DNA using the method of electrophoration (inserting plasmids into?E.coli).Approx. Duration:Prep
DataONE:Protocols/Find－GEO－reuses
Identify reuses of GEO datasetsAimThe aim of this protocol is to collect data on the reuses of datasets in the published literature. This particular p
FISH－protocols－for－Drosophila1
.1 RNA Probe Preparation?(see?Note 1)1.?? 1.5 mL microcentrifuge tubes or standard 96-well V-bottom microplates.2.?? RNAse free water.3.?? T7, T3 or S
Red－Blood－Cell－Lysis－Protocols
實驗概要BioLegend’s ?Red Blood Cell (RBC) Lysis Buffer (Cat. No. 420301) has been designed, ?formulated, and tested to ensure optimal lysis of RBCs in sin
FISH－protocols－for－Drosophila2
?3.?Methods3.1 RNA Probe Preparation1.?? Different strategies can be used to prepare template DNA for synthesizing antisense RNA probes by?in vitro?tr
用DAPI進行細胞染色
用DAPI進行細胞染色DAPI的配置Stock solution of 1mg/ml in ddH2O; working solution of 0.25mg/ml to 50mg/ml in ddH2O or PGM.?1.Place sample on slide.?2.Add a few dr
DAPI的配制及貯存
固體粉末 ：避光，2-8 ℃保存，3年沒有問題。貯存液 ：用無菌水配制成濃度1 mg/ml 的貯存液，配好后用錫紙包起來，避光，可在-20 ℃下長期保存。（DAPI易溶于水，在PBS中溶解度不高）使用濃度 ：貯存液用1xPBS稀釋到終濃度100 ng/ml。熒光封片液 ：0.5 mol/L碳酸鹽緩沖
Streptomyces:Protocols/MiniMaxi－Prep
Small Scale Plasmid Isolation (Mini / Maxi Prep)Description?A mini prep / maxi prep is used to isolate plasmid or cosmid DNA from bacteria, normally E
Standard－Protocols－Autoradiography－(35S)
Remove Kodak NTB2 nuclear emulsion from fridge and place at 42oC for around 30-60 mins (until melted).Make up the developer and the fixer and place in
Cell－Cycle－Staining－ProtocolDAPI
1. Harvest cells- wash 2X in PBS to get rid of serum proteins. 1200rpm, 5 min2. Resuspend pellet (up to 3x106 cells) in 1.2 ml PBS (Ca and Mg free).3.