RestrictionDigestsofHighMolecularWeightYeastDNA
Purpose:To perform restriction digests of YACs for mapping using rare cutting enzymes or more conventional restriction endonuclease digestion.Time required:2-4 hoursSpecial Reagents:dependent upon enzymes desiredProcedure:Using wide-bore pipette, aliquot 100 ng of liquid DNA into eppendorf tube. Add 2.5 祃 10X restriction buffer, 1-2 ul RNase (5 mg/ml stock), 250-500 ng BSA, then dH2O up to 25 祃 volume. Incubate at ap......閱讀全文
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
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
酵母人工染色體
·?????????Easy YAC Preparation Method?(Andrew Davies,Shaw lab)·?????????Screening YAC libraries?(Donis Keller Lab)This is a method for screening YAC l
酵母準備
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
Southen雜交
Southern雜交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
酵母遺傳學技術
Genome-wide Gene Expression Analysis?(Richard Young Research Group,Whitehead Institute for Biomedical Research)A genoe-wide gene expression analysis u
DNA的酶學操作
DNA的酶學操作DNA Modifying Enzymes?(Michael Blaber)Introduction to bacterial restriction/modification system. It provides very useful background knowledge
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
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
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
DNA抽提
DNA抽提（主要內容如下）·???Working with DNA·???DNA Extraction from Bacteria and Other Organisms·???DNA Extraction from Cell and Tissue·???Mitochondria DNA Isola
Denaturing－Gradient－Gel－Electrophoresis－(DGGE)
Purpose:Denaturing gradient gels are used to detect non-RFLP polymorphisms. The small (200-700 bp) genomic restriction fragments are run on a low to h
DNA電泳
DNA電泳（主要內容如下）??Preparation of Agarose Gel and Electrophoresis??Extraction of DNA From Agarose Gel??Extraction of DNA from Acrylamide Gels??DNA Marker?
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
Restriction－Enzyme－Digestion－of－DNA
Materials:10X restriction enzyme buffer (see manufacturer's recommendation)DNAsterile waterrestriction enzymephenol:chloroform (1:1)Add the follow
SQ－Blood－DNA－Midi－Protocol－for－500l3ml－whole－blood
實驗概要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
實驗概要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
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
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
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
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
基因型分析
Randomly Amplified Polymorphic DNA (RAPD)Randomly Amplified Polymorphic DNA (RAPD)?by??(DNA KAFFE)RAPD analysis has been successfully used in mapping
Molecular－Analysis－and－Results－－DNA
Theory of CGHComparative genomic hybridization (CGH) is a fairly new molecular cytogenetic technique that allows detection of DNA sequence copy number
Fungal－Genomic－DNA－Extraction
實驗概要This procedure does not require phenol extraction. The DNA is pure enough for restriction digests, PCR and genomic library construction.High t
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.
蛋白質分子量(protein－molecular－weight)的測定——葡聚糖凝膠過..
實驗原理葡聚糖凝膠（Sephadex）過濾法測定蛋白質分子量的原理，主要是依據這種凝膠具有分子篩作用，一定型號的凝膠具有大體上一定大小的孔徑。在一定的凝膠柱內，凝膠孔隙所占的體積稱為內水容積Vi，凝膠顆粒間的自由空間所占的體積稱為外水容積V0。當樣品流經凝膠柱時，大于孔隙的大分子完全不滲入到凝膠內部
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
Streamlined－DNA－Extraction－Protocol
This method is derived from a procedure developed by Toby Bradshaw and the Poplar Molecular Genetics Cooperative. We have tested the procedure wit