FISHprotocolsforDrosophila2
3. Methods3.1 RNA Probe Preparation1. Different strategies can be used to prepare template DNA for synthesizing antisense RNA probes by in vitro transcription. A gene segment of interest should first be cloned into an appropriate plasmid containing flanking bacteriophage promoter sequences (T3, T7, or Sp6). Then, the plasmid can either be linearized by restriction enzyme digesti......閱讀全文
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
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
細胞遺傳學——原位雜交(ISH)
In Situ Hybridization· ????????In Situ Hybridization?(jsmith1@po-box.mcgill.ca)In situ?hybridization, as the name suggests, is a method of localizing,
IMMUNOHISTOCHEMISTRY－ON－Drosophila－BRAINS
1. Dissect brains in Drosophila Ringers solution.2. Fix 20'' (1hr max) in a 0.5ml microfuge tube with 5 formaldehyde-PBS on ice.3. Rinse 2-3X
Basic－Methods－of－Culturing－Drosophila
實驗概要Basic Methods of Culturing Drosophila實驗步驟Stockkeeping1. Mechanics? ? ? ? Most stocks can be successfully cultured by periodic mass transfer of a
LCM－PROTOCOLS
Slide SectioningParaffin blocks-?For?DNA?analysis:Special LCM processing schedule is followed.The water bath is cleaned using RNAse Zap?, rinsed thoro
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
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,
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
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
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－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
Western－Blotting－Protocols
back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.
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
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
Immunofluorescent－Staining－of－Drosophila－Larval－Brain－Tissue
實驗概要The Drosophila larval brain is a well-established model for investigating the role of stem cells in development. Neuroblasts (neural stem cells)
Immunofluorescent－Staining－of－Drosophila－Larval－Brain－Tissue
INTRODUCTIONThe?Drosophila?larval brain is a well-established model for?investigating the role of stem cells in development. Neuroblasts?(neural stem
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
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
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
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
Live－imaging－with－Drosophila－tissue－culture－cells2
Materials & ReagentsDrosophila?Schneider S2 cellsSchneiders Medium (GIBCO/Invitrogen), 10% fetal calf serum, Antibiotics (Sigma A5955)Depression slide
Live－imaging－with－Drosophila－tissue－culture－cells1
IntroductionLive imaging provides an important complementation to the "snapshot" view obtained in fixed tissue by immunofluorescence. It allows follow
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
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和PRINS技術
一、熒光原位雜交（FISH） 是一種簡單、敏感、而容易操作的方法，結果迅速，并能在同一標本上檢測多個不同的基因，可應用于細胞培養、染色體分裂像、冰凍切片和石蠟切片。 FISH技術是利用特異的DNA探針，標記了生物素，地高辛、或熒光素，對檢測細胞進行DNA-DNA原位雜交，并用熒光法顯示。 FISH