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.) ......阅读全文
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微序列技术
· 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
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
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
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－（二）
DNA preparation by cryotom tissue dissectionPreparations/Materials: Cool cryostat down to -20 to -30°C about 3 hours prior to dissection Label eppendo
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
Western－Blotting－Protocols
back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.
Neutralizing－Bioassay－Protocols
Neutralizing Bioassay ProtocolsIntroductionAntibodies that block binding of cytokines to their specific receptors and neutralize their effects are cri
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－（一）
Metaphase chromosome preparationMaterials: RPMI 1640 medium fetal calf serum (FCS), 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best
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－（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
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
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
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/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
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/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
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
Fluorescence－In－Situ－Hybridization－using－TSA™
实验概要This protocol describes steps for fluorescent in situ hybridization (FISH) to Drosophila embryos using Tyramide Signal Amplification (TSA™), and
Basic－Fluorescent－in－situ－Hybridization－(FISH)
实验概要Fluorescence in situ hybridization method is a kind of physical map drawing method, use fluorescent element mark probe, to detect probe and spli
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