PreparationofYeastDNAEmbeddedinAgarosePlugs
Preparation of Yeast DNA Embedded in Agarose PlugsAnja van Brabant(adapted from Iadonato, S. P., and A. Gnirke. 1996. RARE-cleavage analysis of YACs. Methods Mol Biol 54: 75-85.)1. Inoculate a 5 ml culture with a single colony from a YAC-containing strain of yeast and grow until saturated. Determine the number of yeast cells per milliliter. The cell count should be roughly 1 X 108 cells/ml for a saturated c......阅读全文
Preparation－of－Yeast－DNA－Embedded－in－Agarose－Plugs
Preparation of Yeast DNA Embedded in Agarose PlugsAnja van Brabant(adapted from Iadonato, S. P., and A. Gnirke. 1996. RARE-cleavage analysis of YACs.
酵母准备
Yeast DNA PreparationYeast Genomic Preparation (Gottschling Lab)Rapid method for yeast genomic DNA isolation Yeast DNA Preparation (rapid glass bead
Preparation－of－Agarose－Gels－for－DNA－separations
Weigh out the desired amount of agarose and place in an Erlenmeyer flask with a measured amount of electrophoresis buffer, e.g. for an 0.8% gel, add 0
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
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
High－Molecular－Weight－Yeast－Liquid－DNA－Preparation
Purpose:To isolate intact, high molecular weight DNA from yeast cells for subcloning and rare cutting restriction enzyme analysis. One can expect a yi
Construction－of－BAC－Libraries：Megabase－DNA－Isolation
Megabase DNA IsolationMegabase-size DNA isolation from plantsTo construct large insert DNA libraries in BAC and YAC vectors, methods must be developed
Purifying－Large－E.－coli－Restriction－Fragments－from－PulsedField－Gels
DNA PreparationE. coli chromosomal DNA is prepared following the method of Heath et al. ( J. Bacteriol., 174, 1992). Cells are embedded in agarose, th
Yeast－DNA－Prep
Protocolgrow up yeast culture to appropriate density (near saturation)spin 1.5 mls of culture for 1 min in microfuge and aspirate off supernatantresus
Pulse－Field－Electrophoresis
Manipulating and analyzing DNA are fundamentals in the field of molecular biology. Indeed, separating complex mixtures of DNA into different sized fra
Agarose－Gel－Electrophoresis－of－DNA
1) Dissolve 1 g of agarose in 100 ml of 1X TAE or TBE buffer (gives a 1% gel). See note for making LMP agarose gel. 2) Cast the gel with the comb in p
ELECTROPHORESIS－OF－DNA－IN－AGAROSE－GELS
ELECTROPHORESIS OF DNA IN AGAROSE GELSA). AGAROSE CONCENTRATIONS: Use 0.8% agarose (w/v) for high molecular weight DNA fragments, and 1 - 1.2% f
DNA电泳（agarose胶）
DNA琼脂糖凝胶电泳 实验方法原理 利用DNA分子在琼脂糖凝胶中泳动时具有的电荷效应和分子筛效应。电荷效应是指DNA分子在高于等电点的pH溶液中带负电，在电场
DNA电泳（agarose胶）
DNA电泳可用于：（1）分离不同大小的DNA片段；（2）鉴定目的DNA片段；（3）纯化和回收DNA片段。实验方法原理利用DNA分子在琼脂糖凝胶中泳动时具有的电荷效应和分子筛效应。电荷效应是指DNA分子在高于等电点的pH溶液中带负电，在电场中向正极移动，且相同数量的双链DNA几乎具有等量的净电荷，能以
DNA电泳（agarose胶）
实验材料 DNA样品试剂、试剂盒 琼脂糖 电泳缓冲液溴化乙锭 上样缓冲液仪器、耗材 电泳仪电泳漕 透射紫外灯 胶带纸 紫外成像仪
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
Lambda－DNA－Preparation
Lambda DNA PreparationThis is a plate method that gives very good yield for cloning. I have combined the Promega and Maniatis protocols.Solutions T-TY
细胞遗传学——原位杂交(ISH)
In Situ Hybridization· In Situ Hybridization (jsmith1@po-box.mcgill.ca)In situ hybridization, as the name suggests, is a method of localizing,
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
DNA－Purification－from－Agarose－Gels
1. Separate DNA fragments in an agarose gel cast with 0.5 mg/mL Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel wit
Agarose－Gels－for－Single－Stranded－DNA
1. Prepare 50X TAE as:242 g Tris Base57.1 mL Glacial Acetic Acid100 mL 500 mM EDTA, pH 8.0600 mL ddH2OMix. Bring volume to 1 L. Autoclave.2. Mix the f
Preparation－of－Sonicated－Human－DNA
Purpose:To break up high molecular weight human placental DNA into fragment sizes of 500 bp or less which can be used as competitor DNA in Southern an
Midiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Midiprep Kit allows purification of 100–350 μg of high-quality plasmid DNA from 15–25 mL overnight E. coli cul
Maxiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Maxiprep Kit allows purification of 500–850 μg of high-quality plasmid DNA from 100–200 mL overnight E. coli cu
DNA凝胶电泳（DNA－agarose－gel－electrophoresis）
实验原理琼脂糖凝胶电泳是常用的用于分离、鉴定DNA、RNA分子混合物的方法，这种电泳方法以琼脂凝胶作为支持物，利用DNA分子在泳动时的电荷效应和分子筛效应，达到分离混合物的目的。DNA分子在高于其等电点的溶液中带负电，在电场中向阳极移动。在一定的电场强度下，DNA分子的迁移速度取决于分子筛效应，即分
RNA－analysis－on－nondenaturing－agarose－gel－electrophoresis
实验概要RNA analysis on non-denaturing agarose gel electrophoresis实验步骤1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer
RNA－analysis－on－nondenaturing－agarose－gel－electrophoresis
1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer instead of 1X TBE- use agarose gel in the concentration of 1.1%-1.
Southen杂交
Southern杂交One important thing for transfer:the weight of the object resting on top of the blotting apparatus should not exceed the weight equal to a 5
Quick－Yeast－DNA－Prep:－Isolation－of－Total－DNA－(genomic－and－plasmid)
Grow a 5 ml YPD O/N culture inoculated with a single yeast colony at 30 deg.Transfer culture to a small 13 x 100 glass tube. Spin down cells 2 min. in
Silver:－TimeLapse－Microscopy
Pad Preparation1. Microwave 2% agarose (mix of low-melt and normal, to taste) in Thorn media (see below). (If you have used different percentages of a