CycleSequenceReactionsForLargeInsertPlasmidTemplates
The following dye-labeled terminator reaction chemistries have been designed to balance conservation of reagents with the resulting sequence product signal strength. When coupled with BACPAC Resource template preparation, N-free read lengths of >400nts can be expected. More emphasis can be placed on reagent conservation by further volume reduction or dilution; or on sequence product signal strength by ......閱讀全文
Cycle－Sequence－Reactions－For－Large－Insert－Plasmid－Templates
The following dye-labeled terminator reaction chemistries have been designed to balance conservation of reagents with the resulting sequence product s
DNA測序
DNA測序（主要內容如下）·?????????Sequencing Gel Preparation·?????????Preparation of Templates?·?????????DNA Sequencing by the Dideoxy Method·?????????DNA Sequen
The－reactions－that－feed－amino－groups－into－the－urea－cycle
Excess amino acids in the body can be used as a source of energy, with their carbon skeleton converted to metabolic intermediates such as acetyl-CoA o
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
Protocol－for－dsRNA－Synthesis
實驗概要? ? ? ? We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends
Large－Scale－Plasmid－Preps:－PEG－method
1. Grow 250 - 500 mL of bacteria overnight in LB with 50 μg/mL of ampicillin.2. Transfer culture to Nalgene bottles. Spin in SGA rotor at 6000 rpm, at
Large－Scale－Plasmid－Preps:－Qiagen/Cesium－Method
Most plasmids can be adequately prepped by kits containing DNA binding columns. These columns do not do a great job of separating plasmid DNA from con
質粒的小量制備
·?????????Standard (alkaline lysis) Mini-Prep?(Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
質粒的小量制備
·?????????Standard (alkaline lysis) Mini-Prep?(Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
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
How－do－you－synthesize－your－dsRNA
We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends (I. Primer Desig
重組DNA的分離、克隆與測序實驗手冊8
B. Midiprep double-stranded DNA isolationA midi-prep double-stranded DNA isolation has been developed to generate a sufficient amount of template DNA
Large－Scale－Plasmid,－Cosmid,－BAC,－PAC,－and－Fosmid－DNA－Isolation
DNA Isolation by a Cleared Lysate Method Followed by Double Acetate Precipitation Version 3b - updated September 26, 1999The Most Recent Roe Lab Imple
Quantitative－PCR
實驗概要Quantitative PCR involves co-amplification of two templates: a constant amount of a preparation containing the desired target sequence and var
定量PCR實驗技術－QPCR
Quantitative PCRJoseph SambrookPeter Maccallum Cancer Institute and The University of Melbourne, AustraliaDavid W. RussellUniversity of Texas Southwes
粘粒
Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures?(Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to re
粘粒
Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures?(Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to re
其它PCR方法
·?????????Standard PCR Protocol?(Molecular Biology Techniques Manual)The followings are described in detailRecommended Reagent ConcentrationsRecommend
Ligation－Optimization
The following protocol can be used to optimize ligation conditions for difficult to clone (e.g. very large) fragments. The principle is to independent
Sequencing－off－Cosmid,－BAC,－PAC,－－with－ABI－Big－Dye－Terminators
Big Dye Protocols and Notes - Cosmid, BAC, BAC, Fosmid TemplatesHi all,Over the past two months, we have been testing various reaction conditions for
果蠅RNAi的實驗中雙鏈短RNA的合成（dsRNA)方法
實驗概要We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends (I. Prim
質粒的Midi制備
·?????????Plasmid Midi-preps?(NWSFC)Diatomaceous Earth-based midi-prep. This procedure is the method of choice for isolating double stranded plasmid-b
M13噬菌體
·?????????M13 Phage?(Michael Blaber)Very useful background information about M13: its infection, replication, packing, cloning. If you are new to phag
定量RTPCR－（Quantitative－RTPCR）
Application:?Quantitative RT-PCR?is used to quantify mRNA in both relative and absolute terms. It can be applied for the quantification of mRNA expres
果蠅RNAi的實驗中雙鏈短RNA的合成（dsRNA)方法
本文來自于哈佛大學醫學院果蠅RNAi篩選中心的經典實驗方法，專門用于果蠅RNAi實驗方法。感謝哈佛大學醫學院果蠅RNAi篩選中心的支持！Primer Designed dsRNATemplate SelectionPCRIn vitro?RNA TranscriptiondsRNA Purifica
DNA轉化實驗指導4
2B.??Transformation?1.?????Preparation of electrocompetent DH5a?cells:??autoclave 4 baffled 1 liter flasks containing 500 mL LB.??Remove a 1 mL aliquo
LongPCR－Reagents－and－Guidelines
Long-PCR Reagents and Guidelinesfrom George Church as Modified from Cheng et al. (1)General Guidelines for Long-PCR Conditions and Enzyme Mixtures====
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轉化實驗指導2
1B.??Cloning?1.?????A caveat on dephosphorylation: the most common reason for failure to obtain colonies is a result of adding too much BAP or CIP to
siRNA數據庫與設計工具
siRNA DatabaseSearchable database of Silencer ? Validated and Pre-designed siRNAs to >34,000 human, mouse, and rat targets. All siRNAs in the database