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
HELPER－PHAGE－PREPARATION
HELPER PHAGE PREPARATION1. Grow an overnight of NM522 in NZCYM medium.2. Dilute overnight 1:100 and grow to an A600 = 0.3 (@2.5 x 108 cells/ml).3. Inf
Preparation－of－Phage－Lysates
Preparation of Phage LysatesInoculate 5 ml of lambda-broth in a glass culture tube with a single colony of an appropriate host strain of E. coli. Incu
人工转录因子的部件——人类锌指结构1
Human zinc fingers as building blocks in the construction of artificial transcription factorsKwang-Hee Bae1, 4, Young Do Kwon1, 2, 4, Hyun-Chul Shin1,
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
烈性噬菌体（virulent－phage）和温和噬菌体（temperate－phage）
噬菌体(bacteriaphage or phage)是病毒的一类，结构很简单，基本上由一个蛋白质外壳包裹着一些核酸组成的。噬菌体的多样性来自于组成其外壳的蛋白质的种类，以及其染色体的类型和结构的不同。（一）烈性噬菌体（ virulent phage）遗传学上应用最广泛的烈性噬菌体是大肠杆菌（ E.
DNA抽提
DNA抽提（主要内容如下）· Working with DNA· DNA Extraction from Bacteria and Other Organisms· DNA Extraction from Cell and Tissue· Mitochondria DNA Isola
454高通量测序——研究土壤微生物的新手段（三）
注：已用454焦磷酸测序法发表的文章总结1、Archaea predominate among ammonia-oxidizing prokaryotes in soils—— (2006) Nature 442: 806-809——if 36.1012、454 Pyrosequencing ana
Lambda－Phage－DNA－Quickprep
suspend a single plaque in 1 ml PSBadsorb 10 min at 37°C: 0.1 ml eluted phage*/0.1 ml MgCa/0.1 ml saturated K802 culture grown in NZY broth/0.2% malto
Quantification－made－easy
With the aid of spectroscopy, the quantitative analysis of nucleic acids and proteins has established itself as a routine method in many laboratories.
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
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
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
如何借助epMotion－5073l移液工作站完成微量体积的...（一）
如何借助epMotion 5073l移液工作站完成微量体积的qPCR反应体系构建在进行微量体系的液体操作时，如何避免人为误差与日间变化，如何保证结果的一致性提高实验效率？如果减少冗长枯燥的重复操作，减少昂贵试剂的使用，提升操作体验节省实验经费？本篇应用旨在介绍如何用10 μL分液工具实现全自动的微量
Competitive－RTPCR－Strategy－for－Quantitative－Evaluation－1
Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type IQuantizatio
Preparing－Lambda－DNA
Preparing Lambda DNA1- Coliphage lambda DNA is a widely used vector for recombinant DNA. The middle third of its 48,000 bp contains no genes required
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
Easy－YAC－Preparation－Method
YAC TRANSFORMATION OF C. ELEGANS USING TOTAL YEAST GENOMIC DNA[This method is described in The Worm Breeder's Gazette (1997) A. Davies and J. Shaw
Horizontal－Transfer－of－Supernumerary－Chromosomes－in－Fungi
Several species of filamentous fungi contain so-called dispensable or supernumerary chromosomes. These chromosomes are dispensable for the fungus
Heterogeneity－of－SingleCell－Gene－Expression－Across－Phenotypically（二）
Figure 2: Verification of C1 Single-Cell mRNA-Seq data quality. a)ERCC RNA Spike-In Control Mix 1 was applied to a C1 IFC at a total transcript in
cDNA－Libraries
cDNA LibrariesIsolation of corresponding genetic informationInstead of synthesizing a desired gene, can we used the amino acid information to directly
Differential－cDNA－Screening－Procedures
Differential cDNA Screening ProceduresThe protocols listed refer to cDNA library construction and preliminary differential screening procedures. They
siRNAs结合生物芯片的实验设计2
Figure 2. Silencer ™ siRNA Validation Data Generated Using Applied Biosystems TaqMan® Gene Expression Assays. The indicated Silencer Validated siRNAs
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
Column－Method－for－Lambda－Phage－DNA－Preparation
Purpose:Mini-prep method for lambda phage DNA purification from lysates.Time required:4 hours once the lysate is in handSpecial supplies required:BioR
Organotypic－raft－culture－of－primary－human－keratinocytes－(PHKs)
Organotypic raft culture of primary human keratinocytes (PHKs)NotesUse 106 keratinocytes per raft.Typical ProtocolTrypsinize the PHKs off the flask an
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
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
Sauer:Lysing－E.－coli－with－Lysozymes
Getting The Most Out Of Your BugsNative lysis is a staple protocol in practically every biochemistry lab, yet there is significant variability in the
生物技术等级透析膜应用介绍
At one time or another most scientists working in the ‘biochemical’ sciences will have to dialyze something. Dialysis is a simple technique to exc