HighMolecularWeightYeastLiquidDNAPreparation
Purpose:To isolate intact, high molecular weight DNA from yeast cells for subcloning and rare cutting restriction enzyme analysis. One can expect a yield of 100-200 礸 of DNA per prep.Time required:6 days totalDay 1-3: 10 minutesDay 4: 6-8 hoursDay 5: 2 hoursDay 6: 1 hourSpecial Equipment:23 mm dialysis tubingBeckman SW28 Rotor and UltracentrifugeSpecial Reagents:SCE solutionLyticase (Sigma # L 8137)Yeast lysis buffer......阅读全文
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
Restriction－Digests－of－High－Molecular－Weight－Yeast－DNA
Purpose:To perform restriction digests of YACs for mapping using rare cutting enzymes or more conventional restriction endonuclease digestion.Time req
酵母准备
Yeast DNA PreparationYeast Genomic Preparation (Gottschling Lab)Rapid method for yeast genomic DNA isolation Yeast DNA Preparation (rapid glass bead
DNA－Molecular－Weight－Markers
DNA Molecular Weight Markers Lambda DNA Hind III DigestFragmentBase Pairs123,31029,41536,55744,36152,32262,02775648125Lambda DNA EcoR I DigestFragment
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
酵母遗传学技术
Genome-wide Gene Expression Analysis (Richard Young Research Group,Whitehead Institute for Biomedical Research)A genoe-wide gene expression analysis u
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.
Molecular－Weight－Marker
Molecular Weight Markerl HindIII Digest Marker SolutionDigest 20 µg l DNA (40 µl DNA if at 0.5 µg/µl)Add 50 µl 10X Tracking DyeBring volume of the dig
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
细胞培养——细胞保藏
Working Cell Bank (Contributed by Nanci Donacki)Provides detailed protocol for establishing a working cell bank Master Cell Bank (Contributed by Na
A－practical－method－for－the－preparation－of－total－DNA－from－filamentous－fungi
Most methods of DNA preparation from fungi are time-consuming due to the need to first make protoplasts, expensive for chemicals such as cesium chlori
SQ－Blood－DNA－Midi－Protocol－for－500l3ml－whole－blood
实验概要The E.Z.N.A.® SQ Blood DNA Kit is designed for isolating high molecular weight genomic DNA from fresh, frozen or anticoagulated whole blood. The
SQ－Blood－DNA－Maxi－Protocol－for－410－ml－whole－blood
实验概要The E.Z.N.A.® SQ Blood DNA Kit is designed for isolating high molecular weight genomic DNA from fresh, frozen or anticoagulated whole blood. The
酵母转化
· Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation· Yeast Transformation (Breeden Lab)LiAc method· Large-Scale Y
细胞培养常规操作
常规操作(主要内容如下)· Aseptic Technique· Culture Vessels· Cell Counting· Primary Culture· Maintenance of Cell Line ·
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.
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
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
DNA电泳
DNA电泳（主要内容如下） Preparation of Agarose Gel and Electrophoresis Extraction of DNA From Agarose Gel Extraction of DNA from Acrylamide Gels DNA Marker
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
Preparation－of－High－Titer－Adenovirus－in－P11－cells
adapted from Cell Biology, A Lab Manual, second edition, volume 1 -Grow up P11 cells in 15 cm plates to 70 – 80% confluence. -Infect cells with a MOI
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
巨噬细胞和单核白细胞
· Lymphocyte Transformation (Donis-Keller lab)Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocytes) from antico
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
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
A－safe－method－of－extracting－DNA－from－Coccidioides－immitis
Human-pathogenic fungi such as Coccidioides immitis and Histoplasma capsulatum must be handled in Biosafety level 3 containment facilities which make
质粒的小量制备
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
质粒的小量制备
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
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
Genomic－DNA－Extraction－－PureLink™
实验概要The PureLink™ Genomic DNA Purification Kit allows rapid and efficient purification of genomic DNA. The kit is designed to efficiently isolate g