FusionandCloning
ReagentsMedium A - Pre-fusion Medium and Hybridoma Expansion MediumMedium B - Fusion Medium Medium C - Hybridoma Recovery MediumMedium D - Hybridoma Selection Medium Medium E - Hybridoma Growth Medium PEG SolutionMaterials 50 ml Sterile conical tubes15 ml Sterile conical tubes 10 ml sterile pipets 1 ml sterile pipets Pasteur Pipets, sterile100 mm sterile Petri Dishes 96-well culture dishes24-well culture dishesF......閱讀全文
Fusion－and－Cloning
Author:?Nanci DonackiSource:?Contributed by Nanci DonackiAbstract:?Procedure for establishing hybridoma in one stepReagents(StemCell Technologies, Inc
Fusion－and－Cloning
ReagentsMedium A - Pre-fusion Medium and Hybridoma Expansion MediumMedium B - Fusion Medium Medium C - Hybridoma Recovery MediumMedium D - Hybridoma S
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
Protocol－for－Cell－Fusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, the
Disruption－by－Fusion－PCR
?Disruption by Fusion?PCRDavid Amberg and Ellen Beasley1) In separate PCR reactions, amplify the 5' and 3' ends of the gene of interest with p
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
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
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
Infusion－biobrick－assembly
OverviewThis is a method to assemble two BioBricks using the Clontech In-Fusion PCR Cloning Kit and maintains BioBrick standard formats. There are cur
“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
Cloning－PCR－products－using－TA－vectors
Cloning PCR products using TA vectorsby Paul N. Hengen, Ph.D.?*Methods and reagents is a unique monthly column that highlights current discussions in
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
單克隆抗體
·?????????Tips and hints for the storage of antibodies?(Synaptic Systems)·?????????Purification of IgG Using Protein A- or Protein G-Agarose(KPL)·????
Bacterial－Expression－of－GSTfusion－Proteins
1.? Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture.?2.? Grow larger culture (100x volume of starter culture) using the overnigh
Thrombin－Cleavage－of－GSTFusion－protein
INTRODUCTIONIn many cases the cleavage can be performed using the free intact fusion, or in same cases with the fusion protein bound to a matrix.?The
PC12－Cell－Culture－and－Fusion
Cell CultureMaterials1. Falcon Primaria culture dishes.2. Culture medium: DME (or F12K) with glutamine, supplemented with 7% heat-deactivated horse se
基因克隆技術（Gene－Cloning－Techniques）2
二、目的基因和載體的連接獲得目的基因后必須將其放在一定的載體內才能在宿主細胞內擴增或表達。目的基因與載體的連接及其后續的轉化過程習慣上稱為克隆（cloning）。由于目前很多基因都是利用PCR技術獲得，因此這里先介紹PCR產物的克隆策略，然后再介紹其他的克隆方式。（一）PCR產物的克隆策略獲得PCR
基因克隆技術（Gene－Cloning－Techniques）2
二、目的基因和載體的連接 獲得目的基因后必須將其放在一定的載體內才能在宿主細胞內擴增或表達。目的基因與載體的連接及其后續的轉化過程習慣上稱為克隆（cloning）。由于目前很多基因都是利用PCR技術獲得，因此這里先介紹PCR產物的克隆策略，然后再介紹其他的克隆方式。 （一
Cosmid－Cloning:－Cell－preparation,－DNA－packaging,－and－Cell－Transfection
Cosmid Cloning: Cell preparation, DNA packaging, and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction man
Factor－Xa－Cleavage－of－MBPFusion－protein
INTRODUCTIONIn many cases the cleavage can be performed using the free intact fusion, or in same cases with the fusion protein bound to a matrix.?The
Fusion－Protein－Isolation(融合蛋白分離純化)
Peter Novick Lab,Department of Cell Biology Yale University School of Medicinehttp://info.med.yale.edu/cellbio/Novick/Second/Protocols/Fusion.pdf1.Sta
GST融合蛋白的準備
Preparation of Glutathione-S-Transferase (GST) Fusion ProteinsMargret B. Einarson and Elena N. Pugacheva?Foxx Chase Cancer Center, Philadelphia, PA 19
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends2
3’ Adaptor Ligation and PurificationHeat shock the RNA by putting at 90°C for 30 seconds. Snap cool on ice.Set up the 3’Adaptor ligation reaction in a
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends3
Load your precipitated PCR samples into 2 consecutive lanes so as not to overload the lanes. For each different sample, I would run a separate ladder
Expression－Library－Screening－(Procaryotic)－Using－APFusion－Proteins
Outline:Bacteriophage lambda is a linear double stranded DNA, approximately 50 kB in size. The two sticky ends help the phage to recircularize after e
Experimental－Protocol－for－cDNA－Library－Construction
Experimental Protocol for cDNA Library ConstructionIdentify appropriate celltype over-expressing corresponding gene.Find out if transcription can be s
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends01
IntroductionThe following protocol describes a procedure for the purification and cloning of miRNAs and other small RNAs in the? 20-30 nucleotide size
新品發布－Orbitrap－Fusion－Lumos－Tribrid液相色譜質譜儀
分析測試百科網訊 2015年10月13日，科學服務領域的世界領導者賽默飛世爾科技（以下簡稱：賽默飛）Orbitrap十周年客戶會議在北京北辰洲際酒店隆重召開，在會議期間，賽默飛同時召開了媒體新聞發布會，發布了Orbitrap系列質譜新品—Orbitrap Fusion Lumos Tr
Bacterial－Expression－of－IRS1－containing－GSTfusion－Proteins
1.? Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture.?2.? Grow larger culture using the overnight culture as a seeding culture.?
基因克隆的四大要素(Four－Elements－for－Gene－Cloning)
一、受體細胞 ：受體細胞的選擇是否合適將關系到能否表達，特別是高效表達。首先要注意不同的表達載體與受體細胞的關系，即原核表達載體適合原核細胞，真核載體則適合真核細胞，而農桿菌僅適用于植物細胞。即使大腸桿菌質粒，也應注意選擇合適的菌種。例如，當用含有T7 噬菌體啟動子的載體表達外源基因時，由于T7