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
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
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
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－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
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,
E.coli－Total－RNA－Labeling－Protocol－for－Spotted－Microarray
Note:Start with 20 ug of total RNA for each labeling reaction.All solutions that can be filtered should be filtered.Cy dyes are light sensitive and sh
Standard－Protocols－Autoradiography－(35S)
Remove Kodak NTB2 nuclear emulsion from fridge and place at 42oC for around 30-60 mins (until melted).Make up the developer and the fixer and place in
Streptomyces:Protocols/MiniMaxi－Prep
Small Scale Plasmid Isolation (Mini / Maxi Prep)Description A mini prep / maxi prep is used to isolate plasmid or cosmid DNA from bacteria, normally E
Global－Expression－Profiling－of－RNA－from－Laser－Microdissected－Cells－at－...
Global expression profiling of RNA isolated from laser microdissected cells allows one to profile a specific set of cells allowing for enhanced se
CDNA文库
CDNA文库（主要内容如下）· Construction of cDNA Library· Construction of Genome DNA Library· Library Screening OthersConstruction of cD
DNA－microarray相关的基因表达数据处理攻略
基因表达谱 的发展有助于科研工作人员进一步的理论知识充实及应用到研发等领域中。基因芯片是最近几年发展起来的基因表达重要工具，本文主要对这种技术的数据分析和管理方法作具体介绍。一、引言DNA微阵列(DNA microarray),也叫基因芯片，是近几年发展起来的一种能快速、高效检测DNA片段序列、基因
Microarrays－to－Characterize－the－MolecularGenetic－Basis－of－Disease
article by Dr. RL McInnes, Agilent Technologies, AustraliaGenomic instability underlies cancer and Copy Number Aberrations (CNA s) are known to underp
荧光原位杂交（Fluorescence－in－situ－hybridization,FISH）
实验原理荧光原位杂交（Fluorescence in situ hybridization FISH）是一门新兴的分子细胞遗传学技术，是20世纪80年代末期在原有的放射性原位杂交技术的基础上发展起来的一种非放射性原位杂交技术。目前这项技术已经广泛应用于动植物基因组结构研究、染色体精细结构变异分析、病
Hybridization－of－High－Density－Arrayed－BAC－Nylon－Filter－Blots
Protocol for hybridization of high density arrayed Bacterial Artificial Chromosome nylon filter blots with 100 PCR isolated Unigene cDNA inserts, pool
Slice－and－Explant－Culture－Protocols－–－Hevner－lab－2002
for axon tracing & cell culture studies in vitro using embyros age E11.5 – E16.5modified from Rubenstein lab and Price lab protocols1. Setupa. 2 hr be
Immature－Embryo－Rescue－and－Culture
Embryo culture techniques have many significant applications in plant breeding, as well as basic studies in physiology and biochemistry. Immature
荧光原位杂交（Fluorescence－in－situ－hybridization,FISH）（图）
实验原理荧光原位杂交（Fluorescence in situ hybridization FISH）是一门新兴的分子细胞遗传学技术，是20世纪80年代末期在原有的放射性原位杂交技术的基础上发展起来的一种非放射性原位杂交技术。目前这项技术已经广泛应用于动植物基因组结构研究、染色体精细结构变异分析、病
荧光原位杂交（Fluorescence－in－situ－hybridization,FISH）原理
2）标本变性①将制备好的染色体玻片标本于 50oC培养箱中烤片2～3h。（经Giemsa染色的标本需预先在固定液中退色后再烤片）。②取出玻片标本，将其浸在70～75oC的体积分数70％甲酰胺/2×SSC的变性液中变性2～3min。③立即按顺序将标本经体积分数70％、体积分数90％和体积分数100％冰
双重和多重原位杂交(hybridization－in－situ)技术
为了在同一标本上或同一细胞内同时检测是否存在两种或两种以上的靶核酸序列。可应用双重或多重原位杂交技术．即以两种或多种标记探针与靶核酸杂交。然后利用不同的检测手段分别显示各种靶核酸的存在和分布。该技术与免疫组织化学技术中的双重或多重标记相似，除了探针本身的特异性外，对结果的干扰主要来自标记物及检测试剂
Human－Embryonic－Stem－(ES)－Cell－Protocols——Embryonic－Bodies
Let human ES cells grow until the colonies are large and the cells are pretty piled up - about the time when you would normally split or even a day pa
Human－Embryonic－Stem－(ES)－Cell－Protocols——Media－and－Reagents
Serum Free Media for human ES cells on MEFs: can last for 7-10 daysFinal ConcentrationAmount for 250ml Stock solution80% DMEM-F12200ml20% KO Serum Rep
Multicolour－3DFISH－in－vertebrate－cells3
Pepsin treatmentEquilibrate slides (kept in 50%FA/2xSSC) in 2xSSC 2 minutes;Switch to 1xPBS 3 minutes;Pepsin: 3-5 minutes in 0.01 N HCl/0.002% pepsin.
Protocols－for－the－preparation－of－tumour－cells－for－s.c.－injections－in－mice
For the establishment of solid tumour in nude mice, tumour cells are treated in a sterile environment under a cell culturing hood. Disposable sterile
Protocols－for－the－preparation－of－tumour－cells－for－s.c.－injections－in－mice
For the establishment of solid tumour in nude mice, tumour cells are treated in a sterile environment under a cell culturing hood. Disposable sterile
Human－Embryonic－Stem－(ES)－Cell－Protocols——Matrigel－Aliquoting－and－Plating
Aliquoting Matrigel:Day one:Put the sterilized tip box (either 200 ml or 1000 ml tips), sterilized eppendorf tube container, and appropriate pipettor
Human－Embryonic－Stem－(ES)－Cell－Protocols——Freezing－Human－ES－Cells
Collagenase cells for approximately 7 minutes at 37 °C (until edges of colonies are curling up).With a 5 ml pipet, gently pipet and scrape colonies f
Human－Embryonic－Stem－(ES)－Cell－Protocols——Thawing－Human－ES－cells
Remove Human ES cells from liquid nitrogen storage tank. Fill out a freeze/thaw form.Thaw cryovial by gently swirling in waterbath until only a small
常用分子生物学和细胞生物学实验技术介绍（一）
核酸分子杂交技术由于核酸分子杂交的高度特异性及检测方法的灵敏性，它已成为分子生物学中最常用的基本技术，被广泛应用于基因克隆的筛选，酶切图谱的制作，基因序列的定量和定性分析及基因突变的检测等。其基本原理是具有一定同源性的原条核酸单链在一定的条件下（适宜的温室度及离子强度等）可按碱基互补原成双链。杂交的
Real－Time－PCR－Primer－Sets
Real Time PCR Primer SetsNOW OVER 300 PRIMER SETS!!!UPDATED: NOVEMBER 10th, 2003Quantitative RT-PCR is an important step for the validation of express