DAPICounterstainingProtocols
实验概要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......阅读全文
DAPI－Counterstaining－Protocols
实验概要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
实验概要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特点介绍
DAPI 是一种能够与DNA强力结合的荧光染料。它结合到双链DNA小沟的AT碱基对处，一个DAPI分子可以占据三个碱基对的位置。结合到双链DNA上DAPI分子的荧光强度提高大约20倍，常用与荧光显微镜观测，根据荧光的强度可以确定DNA的量。另外，因为DAPI可以透过完整的细胞膜，它可以用于活细胞和固
什么是DAPI？
DAPI，即4',6-二脒基-2-苯基吲哚，是一种能够与DNA强力结合的荧光染料，常用于荧光显微镜观测。因为DAPI可以透过完整的细胞膜，它可以用于活细胞和固定细胞的染色。
DAPI染色流程
DAPI染色1．原理DAPI为4’，6二脒基-2-苯吲哚(4’，6―diamidino-2―phenylindole)能与双链DNA小槽，特别是AT碱基结合，也可插入少于3个连续AT碱基对的DNA序列中。当它与双链DNA结合时，荧光强度增强20倍，而与单链DNA结合则无荧光增强现象，因此是一种简易、
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
实验概要The procedure presented below describes a method for collecting rat blood.实验步骤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
实验概要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进行细胞染色
用DAPI进行细胞染色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的配制及贮存
固体粉末 ：避光，2-8 ℃保存，3年没有问题。贮存液 ：用无菌水配制成浓度1 mg/ml 的贮存液，配好后用锡纸包起来，避光，可在-20 ℃下长期保存。（DAPI易溶于水，在PBS中溶解度不高）使用浓度 ：贮存液用1xPBS稀释到终浓度100 ng/ml。荧光封片液 ：0.5 mol/L碳酸盐缓冲
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.