SequencingoffCosmid,BAC,PAC,withABIBigDyeTerminators
Big Dye Protocols and Notes - Cosmid, BAC, BAC, Fosmid TemplatesHi all,Over the past two months, we have been testing various reaction conditions for the ABI Big-Dye Terminators with double-stranded DNA templates ranging from pUC or pGEM without any insert up to bacterial genomes in the 2 Mbp size. Below are the results of these experiments and the conclusions we have reached. PLEASE BE ADVISED that any deviation fro......閱讀全文
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
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
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
細菌人工染色體
The Construction of Bacterial Artificial Chromosome (BAC) Libraries (complete manuscript)?(Clemson University Genomics Institute)??Construction of BAC
BAC－EndSequencing
BAC End-Sequencing (Diana Bocskai) For every 4 mls of culture, dissolve the BAC DNA pellet in 40 μl of water. for example: Usually each BAC is g
質粒的大量制備
·?????????Plasmid Mini and Maxi Prep Methods?(Gimila Lab)?·?????????Maxi-preps and all media, solutions?(NWFSC)Isolation of cosmid, plasmid and P1 DNA
質粒的大量制備
·?????????Plasmid Mini and Maxi Prep Methods?(Gimila Lab)?·?????????Maxi-preps and all media, solutions?(NWFSC)Isolation of cosmid, plasmid and P1 DNA
BAC/PAC－文庫的構建
BAC文庫的構建 ? ? ? ? ? ? 實驗方法原理 BAC是一種裝載DNA大片段的克隆載體系統，用于人、動物和植物基因組文庫構建。BAC具有插入片段較大(幾千個堿基至350kb)
BAC/PAC－文庫的構建
BAC (Bacterial Artificial Chromosome，細菌人工染色體)文庫可用于：（1）全基因組測序；（2）構建物理圖譜、染色體步查；（3）基因篩選；（4）基因圖位克隆。實驗方法原理BAC是一種裝載DNA大片段的克隆載體系統，用于人、動物和植物基因組文庫構建。BAC具有插入片段較
DNA－Isolation－From－BAC－－PAC－Clones
DNA Isolation From BAC & PAC Clones? This is a rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modifi
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
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?
鳥槍法如何測序
全基因組鳥槍法測序（Whole Genome Shotgun Sequencing）無需構建各類復雜的物理圖譜和遺傳圖譜，采用最經濟有效基因組鳥槍法測序/鳥槍法Shotgun測序（Whole Genome Shotgun Sequencing）無需構建各類復雜的物理圖譜和遺傳圖譜，采用最經濟有效的實
Template－Preparation
Template PreparationThe quality of sequencing results is directlyrelated to the quality of the template.?ABI recommends a minialkaline-lysis/PEG preci
ABI－3730XL測序平臺的原理及特點
Applied?Biosystems?3730XL測序儀是高質量的長片段讀取和序列分析的測序平臺，應用靈活而廣泛。3730XL可同時分析96個樣品，該儀器采用4色熒光同時檢測，可不間斷24小時運行，自動灌膠，上樣，電泳分離，檢測及數據分析。本儀器除了能夠完成新基因測序工作或比較測序工作外，還可進行多
Mouse－Big－Endothelin,Big－ET－ELISA－Kit使用原理
Mouse Big Endothelin,Big ET ELISA Kit使用原理小鼠大內皮素(BigET)ELISA試劑盒貨號:FK-QZ3291規格：96T/48TMouse Big Endothelin,Big ET ELISA Kit使用原理The kit was a solid pha
ABI－3730XL測序平臺的原理及特點
Applied Biosystems 3730XL測序儀是高質量的長片段讀取和序列分析的測序平臺，應用靈活而廣泛。3730XL可同時分析96個樣品，該儀器采用4色熒光同時檢測，可不間斷24小時運行，自動灌膠，上樣，電泳分離，檢測及數據分析。本儀器除了能夠完成新基因測序工作或比較測序工作外，還可進
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
Cosmid－DNA－Isolation
實驗概要 Isolation of high yields of highly pure cosmid DNA using PureLink? HiPure Plasmid Purification Kits. 實驗原理 The ?PureLink? HiPure Plasmid Purif
Cosmid提取技術
Day OneStart overnight cultures of Cosmid in fresh L-Broth + 25 ug/ml kanamycin. 4 x 250 ml each, inoculate each with metal loop from frozen glycerol
ABI－3730XL測序平臺的原理及特點
Applied Biosystems 3730XL測序儀是高質量的長片段讀取和序列分析的測序平臺，應用靈活而廣泛。3730XL可同時分析96個樣品，該儀器采用4色熒光同時檢測，可不間斷24小時運行，自動灌膠，上樣，電泳分離，檢測及數據分析。本儀器除了能夠完成新基因測序工作或比較測序工作外，還可進
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
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
Plasmid－or－Cosmid－DNA－Miniprep
This protocol can be used to isolate sufficient amount DNA from 1.5ml o/n culture or 3ml 6hr culture to do several enzyme digestions.Spin 1.5ml o/n cu
Construction－of－BAC－Libraries：SOLUTIONS－FOR－BAC－LIBRARY－CONSTRUCTION
SOLUTIONS FOR BAC LIBRARY CONSTRUCTION10X Homogenization Buffer (HB) stock: (1 liter)IngredientAmountFinal ConcentrationTrisma base12.1 g0.1 MKCl59.7
NO－WEDGE－Sequencing－Gels
I have been using an alternative to wedge gels that saves acrylamide, cuts gel drying time to 20 min and gives as good or better band squashing at the
PREPARATION－OF－SEQUENCING－GELS
MATERIALS:2-glass plates1 sharks -tooth comb and spacersWhatman 3 mm paper30 or 40% acrylamide-bis (19:1)10X TBEurea10% ammonium persulfateTEMED60 cc.
DNA－Sequencing－Gels
DNA Sequencing GelsBuffers and gel solutionsLong Ranger: we started using this in early 1995. Great stuff; the best thing is that the gels are not sti
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====