EasyWaytoCloneGenesFromaPhageLibrary
Easy Way to Clone The protocol is oriented towards a C. albicans genomic library I made in Lambda Zap II on 7/97.The overall sequence of events is: • Titer and plate out phage • Lift plaques onto filters and prepare them for screening • Make a probe • Hybridize the probe to the filters • Wash the filters and expose to film • Purify putative plaques • Excise plasmid from the des......阅读全文
Easy－Way－to－Clone－Genes－From－a－Phage－Library
Easy Way to Clone The protocol is oriented towards a C. albicans genomic library I made in Lambda Zap II on 7/97.The overall sequence of events is: •
cDNA－AMPLIFICATION－FROM－LAMBDAPHAGE－LIBRARY
PREPARE SOLUTIONS1. SM buffer (1 L):Mix 5.8 g of NaCl, 2 g of MgSO4-7H2O, 50 mL of 1M Tris-HCl, pH 7.5, 0.5 mL of 2% gelatin, and dH2O to 1 L (Autocla
Preparation－of－phage－particles－from－phage－vectors
Pick up one phage vectors-containing colony with a sterile loop and put into 10 ml 2xTY + 10 µg/l tetracycline.Shake at 200 rpm and 37 °C untill the
Genomic－Libraries
Genomic DNA libraries Size of some genomes and chromosomes:Comparative Sequence Sizes(Bases)(yeast chromosome 3)350 ThousandEscherichia coli (bacteriu
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
Screening－a－cDNA－Library
Screening a cDNA Libraryfor use with HybriZAP zebrafish cDNA librariesObjectivecDNA library screening allows detection of expressed genes for subseque
Phage－Titer
IntroductionLambda phage is a commonly used vector for transgenes. Its very high rate of infectivity makes it ideal for creating large numbers of clon
Quick－and－Easy－Isolation－of－Genomic－DNA－from－Yeast
ProcedureTransfer 1.5 ml of liquid culture of yeast grown for 20 - 24 h at 30°C in YPD (1% yeast extract, 2% peptone, 2% dextrose) into a microcentrif
利用人工组合转录因子对人类基因组扫描2
Figure 5: Regulation of CDH5 by TFZFs in several human cancer cell lines.Blue, cells infected with a pMX construct containing the DNA binding domain o
Degenerate－PCR,－a－short－guide.
What is degenerate PCR? Degenerate PCR is in most respect identical to ordinary PCR, but with one major difference. Instead of using specific PCR pr
Degenerate－PCR
Degenerate PCR is in most respects identical to ordinary PCR, but with one major difference. Instead of using specific PCR primers with a given sequen
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
噬菌体的生长
Preparing Lawn Cells for M13 Cloning (Life Technologies)Lawn cells require the F' episome for M13 infection and may be prepared Streaking Lambda
DNA克隆
DNA克隆（主要内容如下）· General Procedure· PCR Cloning· Subcloning· ET Cloning· Vector Preparation· Ligation Re
The－TRC－shRNA－Design－Methods－and－Rules
OverviewWe design shRNA molecules with an algorithm. Our algorithm uses several criteria to rank potential 21mer targets within each human and mouse R
Construction－and－Manipulation－of－LargeInsert－Bacterial－Clone－Libraries
Acknowledgements The organizer of the workshop acknowledges Dr. Murray Milford, Professor and Interim Head, and Dr. Mark Hussey, Professor and Interim
Standard－Operating－Procedures－for－T1Phage－Testing－Assay
I. Introduction:This assay uses a lawn of phage-susceptible E. coli (DH10B) embedded in a layer of agarose. This top agarose lays on a bed of standard
人工转录因子的部件——人类锌指结构2
Table 2: Binding sites and identity of ZFPs used in VEGF activationWe then generated artificial transcription factors by fusing the three-finger domai
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
The－TRC－shRNA设计方法与原则
OverviewWe design shRNA molecules with an algorithm. Our algorithm uses several criteria to rank potential 21mer targets within each human and mouse R
Twohybrid－analysis－of－genetic－regulatory－networks2
2.2 Interaction mating - large scaleWith a few modifications, the procedure described above can be used to test for interactions between a single prey
Microaspiration－of－Esophageal－Gland－Cells－and－cDNA－Library－Construction－for
Microaspiration of Esophageal Gland Cells and cDNA Library Construction for Identifying Parasitism Genes of Plant-Parasitic NematodesIdentifying p
M13噬菌体
· M13 Phage (Michael Blaber)Very useful background information about M13: its infection, replication, packing, cloning. If you are new to phag
Phage－DNA
IntroductionThe phage lysate from the plate contains bacterial DNA and RNA, as well as phage DNA encased in the phage coat. The following procedure, d
sothing－about－Genome－walking
Can anyone recommend a good kit for genome walking? I would like to find out the promoter sequence of a known gene. Is genome walking the way to do it
DNA的酶学操作
DNA的酶学操作DNA Modifying Enzymes (Michael Blaber)Introduction to bacterial restriction/modification system. It provides very useful background knowledge
Performing－a－hunt－by－interaction－mating
AbstractWhen more than one bait will be used to screen a single library, significant time and resources can be saved by performing the interactor hunt
组织学——显微解剖
Laser Capture Microdissection (LCM)Introduction to LCM (BJMU) Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections (NIH Laser Capture M
利用人工组合转录因子对人类基因组扫描
Scanning the human genome with combinatorial transcription factor librariesPublished online: 18 February 2003, doi:10.1038/nbt794March 2003 Volume 21
酵母人工染色体
· Easy YAC Preparation Method (Andrew Davies,Shaw lab)· Screening YAC libraries (Donis Keller Lab)This is a method for screening YAC l