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
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、
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－（四）
CGH Image acquisitionImages were acquired through a Zeiss Axiophot fluorescence microscope using a Plan NEOFLUAR oil objective x63, N.A. 1.25 (Zeiss,
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
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
CGH－Protocols－（一）
Metaphase chromosome preparationMaterials: RPMI 1640 medium fetal calf serum (FCS), 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best
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－（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
Western－Blotting－Protocols
back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.
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
Neutralizing－Bioassay－Protocols
Neutralizing Bioassay ProtocolsIntroductionAntibodies that block binding of cytokines to their specific receptors and neutralize their effects are cri
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
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 #
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
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
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
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
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－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
DAPI染色法的步骤
原来用dapi是用做原位杂交的配置的时候用灭菌水配置就可以了，以前用的浓度时1ml里面加2ul就可以了配置好的溶液保存在4℃就行，尽量避光，原液要用黑色的或锡箔纸包好放到-20℃为好原来是染载玻片上的染色体，直接滴到载玻片上就好了，之后用缓冲液洗净注意事项就是别沾到手上了，那个东西还是很毒的，染色的
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.
DAPI的属性介绍荧光属性
当DAPI与双股DNA结合时,在荧光显微镜观察下，DAPI染剂在358nm(紫外线)处有一个最大吸收峰，并在461nm(蓝)处有一个最大发射峰，其发射光的波长范围含盖了蓝色至青绿色，因此在荧光显微技术中DAPI被紫外线照亮且被蓝或青色滤镜检出。DAPI也可以和RNA结合，但产生的荧光强度不及与DNA