GeneralCloningProtocols
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. with a single colony. Use all 5 mL to inoculate a 500 mL LB/Amp culture in the evening. Alternatively the 5 mL culture can also be set up as an overnight culture.Save 1 mL for a glycerol stock if necessary (see step 6c, below). Prepare remainder according to ......閱讀全文
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
Human－Embryonic－Stem－(ES)－Cell－Protocols——General－notes－on－ES－cell－culture
hES media has a two week shelf life.hES cells should be cultured in 4, 6, 24, 48, 96 well plates. Growing cells in flasks is not recommended because i
Targeted－Gene－Replacement－in－Fungal－Pathogens－via－Agrobacterium－...
Genome sequence data on fungal pathogens provide the opportunity to carry out a reverse genetics approach to uncover gene function. Efficient meth
General－Reference
Units1 mg = 10-3 g1 ug = 10-6 g1 ng = 10-9 g1 pg = 10-12 g1 kb of double stranded DNA = 660 kD?1 kb of single stranded DNA = 330 kD?1 kb of single str
Bacterial－transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
Fusion－and－Cloning
ReagentsMedium A - Pre-fusion Medium and Hybridoma Expansion MediumMedium B - Fusion Medium Medium C - Hybridoma Recovery MediumMedium D - Hybridoma S
Fusion－and－Cloning
Author:?Nanci DonackiSource:?Contributed by Nanci DonackiAbstract:?Procedure for establishing hybridoma in one stepReagents(StemCell Technologies, Inc
LCM－PROTOCOLS
Slide SectioningParaffin blocks-?For?DNA?analysis:Special LCM processing schedule is followed.The water bath is cleaned using RNAse Zap?, rinsed thoro
DNA克隆
DNA克隆（主要內容如下）·?????????General Procedure·?????????PCR Cloning·?????????Subcloning·?????????ET Cloning·?????????Vector Preparation·?????????Ligation Re
immunofluorescence－of－general－PBMC－by－Peprotech
實驗概要The following protocol provides a method of immunofluorescence of general PBMC by Peprotech.實驗步驟The following protocol used human PBMC that were
immunofluorescence－of－general－cell－by－Peprotech
實驗概要The following protocol provides a method of immunofluorescence of general cell by Peprotech.實驗步驟Please refer to the antibody Product Information S
General－Laboratory－Methods－for－Tetrapyrroles
There are thousands of porphyrins and metalloporphyrins, and hundreds of new derivatives appear each year. This variety arises because the cyclic
ELISA－Protocol－(General－Guidelines)
實驗概要Sandwich ?enzyme-linked immunosorbent assays (ELISAs) involve attachment of a ?capture antibody to a solid phase support. Samples containing known
DNA－EXTRACTION－PROCEDURE－－GENERAL
Grow cells overnight in 500 ml broth medium.Pellet cells by centrifugation, and resuspend in 5 ml 50 mM Tris (pH 8.0), 50 mM EDTA.Freeze cell suspensi
ELISA－Protocol－(General－Guidelines)
實驗概要Sandwich ?enzyme-linked immunosorbent assays (ELISAs) involve attachment of a ?capture antibody to a solid phase support. Samples containing known
Genomic－Cloning－Technical－Manual
Genomic Cloning Technical ManualAn optimal strategy for genomic cloning should meet three requirements: 1) a maximum number of recombinants should be
基因cloning經驗指南
我們克隆基因的時候，往往可以通過一些途徑（如pcr,EST庫或者文庫篩選）得到基因的部分片段。然后通過3'race 和 5'race 方法往兩端延伸。有時會出現無法延伸的狀況，如果你超作沒有失誤的話，這時候很有可能是你模板GC含量過高的緣故，普通pcr 是沒有辦法延伸的，可以用擴高g
Cloning－by－Limiting－Dilution－of－Hybridoma
Author:?Nanci DonackiSource:?Contributed by Nanci DonackiDate Added:?Tue May 14 2002Date Modified:?Tue Apr 27 2004MaterialsDMEM, high glucose (Life Te
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.
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－（四）
CGH Image acquisitionImages were acquired through a Zeiss Axiophot fluorescence microscope using a Plan NEOFLUAR oil objective x63, N.A. 1.25 (Zeiss,
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
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/PCR
Description?Polymerase Chain Reaction (PCR) is a method of amplifying a specific DNA target sequence. The cycle involves denaturing the template doubl
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
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
“電子”基因克隆－（sillcon－cloning）
利用計算機來協助克隆 基因，稱為“電子”基因克隆 （sillcon cloning），是與定位克隆 、定位候選克隆 策略并列的方法之一，即采用生物信息學的方法延伸EST序列，以獲得基因部分乃至全長的cDNA序列。EST數據庫的迅速擴張，已經并將繼續導致識別與克隆 新基因策略發生革命性變化。1
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
