ORNLMICROARRAYHYBRIDIZATIONPROTOCOLS
Direct labeling of total RNA with Cy3 and Cy5:A. MATERIALSRNeasy? Mini Kit (Qiagen; Cat # 74106) SuperScript II RT (200U/μL) (Life Technologies; Cat # 18064-014) QIAquick PCR Purification Kit (Qiagen; Cat # 28106) 100 mM dNTP Set PCR grade (Life Technologies; Cat # 10297-018) Primers: Poly d(T) [20-mers] 2.5 mg/mL (avoids labeling the gDNA and rRNA.) ......閱讀全文
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 #
Microarray－Hybridization－Protocol
Introduction:One microarray set consists of 7 nylon membranes with 2.5 x 7.5 cm dimension. 2304 genes were spotted onto nylon membranes (Schleicher an
DNA微序列技術
·?????????Protocols for Making Drosophila Arrays?(Stanford U.)Detailed protocol for making arrays including PCR Amplification of cDNAs for Printing,?
GenomeWide－Identification－of－Transcription－FactorBinding－Sites－in...
Genome-Wide Identification of Transcription Factor-Binding Sites in Plants Using Chromatin Immunoprecipitation Followed by Microarray (ChIP-chip)
RNA－isolation－for－Microarray
Description?RNA extraction using TRI REAGENT. This method gives ample amout of RNA.Procedure?It is 3 days procedure.Day 1:1. Harvest the cells and cen
Colony－Hybridization
ProcedurePrepare serial 10-fold dilutions of transformed bacteria in LB and spread 100 μL onto LB/Amp plates, as described in the Electroporation prot
In－situ－hybridization
Note:?Although it is possible to perform in situ hybridization on bleached embryos, it appears to reduced the strength of the signal. For best results
COLONY－HYBRIDIZATION
COLONY HYBRIDIZATION1) CUT 4 PIECES OF 3MM WHATMAN PAPER AND PLACE EACH ONE IN A SEPARATE CONTAINER(A CAFETERIA TRAY WILL PROBABLY WORK PERFECTLY).2)
LCM－PROTOCOLS
Slide SectioningParaffin blocks-?For?DNA?analysis:Special LCM processing schedule is followed.The water bath is cleaned using RNAse Zap?, rinsed thoro
Principles－of－nucleic－acid－hybridization
Principles of nucleic acid hybridization5.2.1.?Nucleic acid hybridization is a method for identifying closely related nucleic acid molecules within tw
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
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,
Western－Blotting－Protocols
back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.
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
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
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
ChIPChip－E.－coli
AbstractChIP-Chip stands for?Chromatin?Immunoprecipitation and chip in the sense of DNA microarray. It is a technique to determine the genome-wide bin
Basic－Fluorescent－in－situ－Hybridization－(FISH)
實驗概要Fluorescence ?in situ hybridization method is a kind of physical map drawing method, ?use fluorescent element mark probe, to detect probe and spli
Making－RNA－probes－for－in－situ－hybridization
Make DNA templates via PCRDay 1It's preferable to start with constructs that contain RNA polymerase start sites (T3, T7, or SP6) which allow use o
In－Situ－Hybridization－to－Somatic－Chromosomes－in－Drosophila
In Situ Hybridization to Somatic Chromosomes in DrosophilaAbby F. DernburgINTRODUCTIONIn situ hybridization was originally developed as a technique fo
Fluorescence－In－Situ－Hybridization－using－TSA?
實驗概要This ?protocol describes steps for fluorescent in situ hybridization (FISH) ?to Drosophila embryos using Tyramide Signal Amplification (TSA?), and
RNA－FISH－on－cultured－cells－in－interphase
IntroductionFluorescence?in situ?hybridization (FISH) has become a widely used method in genome and molecular genetic studies. The technique is highly
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
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