FISHprotocolsforDrosophila1
.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 SP6 RNA Polymerase (Fermentas Life Sciences, Burlington, ON, Canada; Catalog Nos. EP0101, EP0111, EP0131) as appropriate.4. 10x transcription buffer (supplied with polymerases: 0.4 M Tris-HCl, pH 8.0; 60 mM MgCl2, 100 m......阅读全文
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
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
细胞遗传学——原位杂交(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
实验概要Basic Methods of Culturing Drosophila实验步骤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
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
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/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
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,
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
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
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
Western－Blotting－Protocols
back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.
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
Immunofluorescent－Staining－of－Drosophila－Larval－Brain－Tissue
实验概要The Drosophila larval brain is a well-established model for investigating the role of stem cells in development. Neuroblasts (neural stem cells)
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
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
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 #
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
Live－imaging－with－Drosophila－tissue－culture－cells2
Materials & ReagentsDrosophila Schneider S2 cellsSchneiders Medium (GIBCO/Invitrogen), 10% fetal calf serum, Antibiotics (Sigma A5955)Depression slide
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
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－技术基本实验
实验方法原理 试剂、试剂盒 70%乙醇SSC磷酸盐缓冲溶液（PBS)Hemo-De清理试剂蛋白酶溶液仪器、耗材 水浴锅增湿器玻片实验步骤 一、制备培养的中期细胞标本1.在一个独立小室里放置一个水浴锅和一台增湿器。湿度应大约50%，如果房间湿度计显示低于45%，则应使用加湿器。2.预热水浴锅至67℃±