MicroarrayHybridizationProtocol
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 and Schuell GmbH, Dassel, Germany) using a GMS417 Microarrayer (Genetic Microsystems, Woburn, MA) at 665 ?m distance between spots.Preparation of the cDNA probe (one microarray membrane set)In tube A prepare: 100mg Total RNADEPC water to 100 ?lIncubate the tube at 70......阅读全文
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
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 #
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
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,
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
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
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)
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
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
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
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,
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
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
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™
实验概要This protocol describes steps for fluorescent in situ hybridization (FISH) to Drosophila embryos using Tyramide Signal Amplification (TSA™), and
Screening－BAC－filters－with－nonradioactive－probes
(Protocol for a single high-density BAC colony filter (HDR filter) of 22 cmx22 cm) to be screened using Amersham''s ECL hybridization kit; thi
Genomic－Southern－Blot－Analysis
This chapter describes a detailed protocol for genomic Southern blot analysis which can be used to detect transgene or endogenous gene sequences i
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
cDNA
· cDNA Synthesis (Crawford Lab)mRNA can be converted into DNA (copy DNA, cDNA) by annealing oligo-dT to the 3' poly-A tail that occurs on
细胞遗传学——比较基因组杂交(CGH)
· Comparative Genomic Hybridization (CGH) CGH is a molecular Cytogenetic method of screening a tumor for genetic changes. The alterations are
NAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
Immunoblot－Protocol
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
ELISPOT－Protocol
实验概要The Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method of measuring the antibody or cytokine production of immune cells on t
ELISPOT－Protocol
实验概要The Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method of measuring the antibody or cytokine production of immune cells on t
RLGS－protocol
A. Preparation of DNA SolutionIn the case of rice, for example This method may be appllicable for many grass species and some other plants.
RNAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
PCR－protocol
PCR reactionProtocol for 50µl reaction - adjust amounts if necessary, for a 20µl reaction use the same volumes of primer and dNTP-mix, but adjust the
ELISPOT－protocol
实验概要The procedure below is a general guideline procedure for ELISPOT. Abcam ELISPOT kits have been designed for detection of various cytokines and g