人VL基因文庫（genomiclibrary）的構建
[器材和試劑] ● PCR試劑和設備 ● cFv基因文庫單鏈模板DNA,制備自pHENl中的天然scFv文庫（10ng/u1） ● Gelleclean試劑盒（Qbiogene） ● Wtzard PCR純化試劑盒（ProlneSa） ● RJHl/2Xho引物： 5'-GGC ACC CTG GTC ACC GTC TCG AGT GGT GGA-3' ● RJH3Xho引物： 5'-GGG ACA ATG GTC ACC GTC TCG AGT GGT GGA-3'● RJH4/5Xho引物： 5'-GGA ACC CTG GTC ACC GTC TCG AGT GGT GGA-3' ● RJH6Xho引物： 5'-GGG ACC ACG GTC ACC GTC TCG AGT GGT GGA-3&#......閱讀全文
人VL基因文庫（genomic－library）的構建
[器材和試劑]?● PCR試劑和設備?● cFv基因文庫單鏈模板DNA,制備自pHENl中的天然scFv文庫（10ng/u1）?● Gelleclean試劑盒（Qbiogene）?● Wtzard PCR純化試劑盒（ProlneSa）?● RJHl/2Xho引物： 5'-GGC ACC CT
人VL基因文庫的構建
[器材和試劑]●? PCR試劑和設備●? cFv基因文庫單鏈模板DNA,制備自pHENl中的天然scFv文庫（10ng/u1）●? Gelleclean試劑盒（Qbiogene）●? Wtzard PCR純化試劑盒（ProlneSa）●? RJHl/2Xho引物：? 5'-GGC ACC C
PCR剪接VHCDR3基因文庫和VL基因構建scFv基因文庫
[器材和試劑]Winard PCR純化試劑盒 （Promega）PCR試劑和設備用于連接scFv和pHENl DNA以及將scFv文庫電轉化到大腸桿菌TGl株的試劑和設備 FDSEQ引物擴增的VHCDR3基因文庫和VL基因，[方法]1. 配制4個25ulPCR反應液，包含：去離子水，10.25ul1
PCR剪接VHCDR3基因文庫和VL基因構建scFv基因文庫
[器材和試劑]Winard?PCR純化試劑盒?（Promega）PCR試劑和設備用于連接scFv和pHENl?DNA以及將scFv文庫電轉化到大腸桿菌TGl株的試劑和設備 FDSEQ引物擴增的VHCDR3基因文庫和VL基因，[方法]1. 配制4個25ulPCR反應液，包含：去離子水，10.25ul1
VHCDR3基因文庫與VL基因的擴增
[器材和試劑]● PCR試劑和設備● Geneclean試劑盒（Qbiogene）● VHFOR隨機引物（見下文），10pmol/ul● LMB3引物● VHBACK引物； 5,-TTT GAC TAC TGG GGCCAG GG-3', 10pmol/u1● FdSEQ 引物： 5'
NGS基因文庫構建大比拼（二）
隨著測序技術的發展，科學界也開始越來越多地應用測序技術來解決生物學問題。在過去的十年里，新一代測序技術—第二代測序(NGS)迅猛發展，越來越多測序實驗室的采用NGS技術作為測序主要手段。而且NGS高通量的特點，使之成為研究DNA和RNA的主要工具。在NGS過程中，構建基因文庫是整個測序進程中第一個步
抗原抗體反應抗體基因文庫
抗體基因文庫（antibody recombination library）是將不同的重鏈和輕鏈基因隨機組合，克隆到合適的表達載體中，在原核細胞表達不同的抗體，形成一個抗體庫，從這個抗體庫中，用抗原可以篩選到相應的抗體基因。抗體基因來源于雜交瘤細胞或動物B細胞（免疫或未免疫）的DNA和mRNA。
用突變的VlCDR3構建SCFv基因文庫
實驗概要本實驗用突變的VlCDR3構建了SCFv基因文庫。主要試劑1. 去離子水(Millipore)2. VentDNA聚合酶和緩沖液(NEB)3. 20XdNTP(每種濃度5mmol/L)(NEB)4. 瓊脂糖膠盒5. Geneclean試劑盒(QbiOgene)6. 含有起始scFv基因的質粒
Genomic－Libraries
Genomic DNA libraries?Size of some genomes and chromosomes:Comparative Sequence Sizes(Bases)(yeast chromosome 3)350 ThousandEscherichia coli (bacteriu
Library－cDNA－Synthesis
Library cDNA Synthesis1° cDNA SynthesisN.B:?During 1° cDNA synthesis, all steps should be carried while wearing gloves and all solutions should either
Screening－a－cDNA－Library
Screening a cDNA Libraryfor use with HybriZAP zebrafish cDNA librariesObjectivecDNA library screening allows detection of expressed genes for subseque
cDNA－LIBRARY－SCREENING
PREPARE SOLUTIONS1. 10mM MgSO4, 0.2% Maltose LB (100 mL):Mix?1.0?g of Bacto-Tryptone,?1.0?g of NaCl,?0.5?g of Yeast Extract, and?1.0?mL of 1M MgSO4. A
Genomic－Southern－Blot
SolutionsProtocol:Digest 5-10 μg genomic DNA overnight with restriction enzyme of choice.Run digested gDNA on 0.8% TAE gel with marker (with no ethidi
Organelle－DNA－Library－Construction
Organelle DNA Library Construction(version MAY-1998)I.?NEBULIZATION?of?DNA ?????1.?0.5?-?5?ug?DNA?in?TE?(10mM/1mM),?25%?glycerol,?final?volume?500?ul
How－to－build－a－BAC－library
Introduction???The?most?important?aspect??of?our?cloning??vectors?is?that?they?are based?on???the?E.?coli?F-factor???replicon.?It?allows?for??strict?
什么是噬菌體肽文庫？
中文名稱噬菌體肽文庫英文名稱phage peptide library定 義將編碼多肽的外源基因插入含噬菌體外殼蛋白基因的載體，構建得到能與外殼蛋白融合表達多肽的基因文庫。應用學科細胞生物學（一級學科），細胞生物學技術（二級學科）
噬菌體肽文庫的定義
中文名稱噬菌體肽文庫英文名稱phage peptide library定 義將編碼多肽的外源基因插入含噬菌體外殼蛋白基因的載體，構建得到能與外殼蛋白融合表達多肽的基因文庫。應用學科細胞生物學（一級學科），細胞生物學技術（二級學科）
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
Fungal－Genomic－DNA－Extraction
實驗概要This procedure does not require phenol extraction. The DNA is pure enough for restriction digests, PCR and genomic library construction.High t
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
Yeast－Genomic－DNA－Prep
Grow 10ml YPD cultures o/n. Figure out cell density; inoculate 30 ml YPD and grow o/n so that cell density is approximately 2 x 108cells/ml the next m
Fungal－Genomic－DNA－Extraction
OverviewHigh throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes, half full with liquid
Genomic－DNA－Extraction－－PureLink?
實驗概要The ?PureLink? Genomic DNA Purification Kit allows rapid and efficient ?purification of genomic DNA. The kit is designed to efficiently isolate ?g
Automated－Genomic－DNA－Extraction
實驗概要This section ?provides a general protocol for automated isolation of genomic DNA from ?10-20 μl blood samples in a 96-well format using the Charge
Fungal－Genomic－DNA－Extraction
OverviewHigh throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes, half full with liquid
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
CDNA文庫
?CDNA文庫（主要內容如下）·?????????Construction of cDNA Library·?????????Construction of Genome DNA Library·?????????Library Screening??OthersConstruction of cD
Construction－of－BAC－Libraries：Construction－of－a－BAC－library
Once high molecular weight (HMW) DNA has been prepared it must somehow be fragmented and DNA in the desired size range isolated. In general, as the de
Using－GenBank－for－Genomic－Authentication:－A－Tutorial
The GenBank? database is perhaps one of the most important repositories of genetic information. A researcher working in the field of genomic authe
Genomic－DNA－Extraction－－Phenol－|－Chloroform
實驗概要This section provides a general protocol for genomic DNA extraction using phenol and chloroform.主要試劑1.?????? Glycogen (20 μg/μL)2.?????? 7.5 M NH4