其他模型

网络 第一章　细胞和组织的损伤 第一节　概述 　　生活机体的细胞和组织经常不断地接受内外环境各种刺激因子的影响，并通过自身的反应和调节机制对刺激作出应答反应。这种反应能力可保证细胞和组织的正常功能，维护细胞、器官乃至整个机体的生存。但细胞和组织并非能适应所有刺激的影响，当刺激的性质、强度和持续时间超越了一定的界限时，细胞乃受损甚至死亡。 　　然 ...

网络 绪论 　　疾病是一个极其复杂的过程。在病原因子和机体反应功能的相互作用下，患病机体有关部分的形态结构、代谢和功能都会发生种种改变，这是研究和认识疾病的重要依据。病理学（patho1ogy）的任务就是运用各种方法研究疾病的原因（病因学，ethiology）、在病因作用下疾病发生发展的过程（发病学，pathogenesis）以及机体在疾病过程中的功能、代 ...

DNA Preparation for microinjection or electroporation Pronuclear injection to produce transgenic mice Morula Aggregation to produce chimeric mice Transgenic identificationPCR of DNA from ear clip tis ...

image of a whole mount from a 4 wk old virgin ProtocolSpread tissue on glass slideFix in Carnoy's fixative for 2 to 4 hours at r.t.Wash in 70 % EtOH for 15 minChange gradually to distilled waterRinse ...

Wholemount in situ hybridisationBased on Wilkinson protocol modified by Murray Hargrave (m.hargrave@cmcb.uq.edu.au) Koopman labA. SYNTHESIS OF PROBE1 Mix in the following order at room temperature:ste ...

BAC DNA has been purified successfully for microinjection by three principle methods: Anion Exchange Chromatography (Nucleobond Columns) CsCl Gradient or Sepharose 4B-CL Chromatography. Any of these m ...

This method was established in the University of Michigan Transgenic Core for our users. The method is used to purify BAC DNA for microinjection into fertilized mouse eggs and BAC transgenic mouse pro ...

Alkaline Lysis and CsCl Purification of BAC DNA for Transgenic Production.This method was contributed by Michelle Southard-Smith and Ron Chandler at Vanderbilt University Nashville Tennessee. This pro ...

Microinjection DNA PurificationWe have found the NucleoSpinTM Extract Kit (available from Clontech Catalog number K3051-1) is a simple and fast way to obtain microinjection quality DNA. This is not to ...

DNA ultrapurificationThe success of DNA purification can have profound consequences on the success or failure of any injection effort. This summary of techniques from several different sources was com ...

Isolation of DNA from Mouse Tail Biopsies1. Obtain tail biopsies from 2 to 3 week old mice: Hold mouse firmly at base of tail with one hand with the other cut off 0.5 to 1.0 cm of the tail tip with a ...

LacZ-cell Staining Materials: 1) 0.5% glutaraldehyde in PBS 2) 20mM K4Fe(CN)6o3H2O 0.44g into 50 ml PBS 3) 20mM K3Fe(CN)6 0.33g into 50 ml PBS 4) X-gal 2% in DMF 5) PBS containing 1mM MgCl2 6) Stainin ...

The purity of the transgene DNA used for microinjection is critical for the successful production of transgenic founder mice. DNA impurities in the form of bacterial endotoxins or organic contaminant ...

Mouse Tail (or organ) Biopsy DNA ExtractionLysis Buffer Proteinase K25 ml 1M Tris pH 8.0 (FC 50mM) 100 mg dry Proteinase K (Sigma #2308)50 ml 0.25M EDTA (FC 25 mM) 4.7 ...

1. Obtain the last 2mm of tail and place directly into 200ul 1X PCR Bufferwith Nonionic Detergents (PBND) in a 1.5ml microfuge tube. (Tails can bestored at frozen in PBS or PBND until use.)2. Add 1.2 ...

Sepharose 4B-CL Chromatography BAC DNA Purification for Transgenic ProductionThis method was contributed by Wes Dunnick at the University of Michigan Medical School in Ann Arbor Michigan. The approac ...

Purification of Gene Targeting Vector DNA for Electroporation1. Purify plasmid from bacteria.We recommend the Qiagen EndoFree Plasmid Maxi kit for the purification of the targeting vector plasmid from ...

Sample Preparation for Microinjection General CommentsMicroinjection is the loading or transfer of a dissolved substance into a living cell. The microscopic tip of the glass microcapillary has an inne ...

Use 4ml polycarbonate snap-cap tubes for all steps. Rocking of tubes should be performed at all times. All steps are carried out at room temperature unless otherwise stated.Day 01. Dissect embryos in ...

X-gal staining of embryosProtocol supplied by S. P. Yee (spyee@julian.uwo.ca) and Joe Chan (cchan@hgmp.mrc.ac.uk).ProcedureDissect embryos in PBS Immediately transfer the embryos to fixative at 4oC i ...