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xiaoxiao银虫 (小有名气)
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分子生物学常用实验技术介绍与指南大全，经典书籍下载
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一、《Molecular Cloning 3》 英文版，主要介绍分子克隆的基本实验技术操作，新手如果看不明白此版，可以自己买一本军科院翻译的中文版。 Table of Contents Chapter 1: Plasmids and Their Usefulness in Molecular Cloning Chapter 2: Bacteriophage and Its Vectors Chapter 3: Working with Bacteriophage M13 Vectors Chapter 4: Working with High-Capacity Vectors Chapter 5: Gel Electrophoresis of DNA and Pulsed-Field Agarose Chapter 6: Preparation and Analysis of Eukaryotic Genomic DNA Chapter 7: Extraction, Purification, and Analysis of mRNA from Eukaryotic Cells Chapter 8: In Vitro Amplification of DNA by the Polymerase Chain Reaction Chapter 9: Preparation of Radiolabeled DNA and RNA Probes Chapter 10: Working with Synthetic Oligonucleotide Probes Chapter 11: Preparation of cDNA Libraries and Gene Identification Chapter 12: DNA Sequencing Chapter 13: Mutagenesis Chapter 14: Screening Expression Libraries Chapter 15: Expression of Cloned Genes in Escherichia coli Chapter 16: Introducing Cloned Genes into Cultured Mammalian Cells Chapter 17: Analysis of Gene Expression in Cultured Mammalian Cells Chapter 18: Protein Interaction Technologies http://mededucation.bjmu.edu.cn/mc3/molecular%20clonging%203.pdf 二、Subcloning Notebook Subcloning is a basic molecular biology procedure used to move inserts from one vector to another. Essentially all subcloning reactions proceed as follows: You release and purify your insert from the parent vector, ligate the insert into a prepared destination vector, transform the ligation reaction into competent bacterial cells, then screen the transformed cells for the insert. The Subcloning Notebook will lead you through every step in this process. Table of Contents Page Chapter 1: Classic Subcloning (.pdf, 845kb) Basic Steps for Subcloning Subcloning Strategy Restriction Digestion Double Enzyme Digests Partial Restriction Digestion Creating Blunt Ends Dephosphorylating Vectors Ligation Purifying Vector and Insert Gel Electrophoresis DNA Markers Ordering Information 3 Chapter 2: PCR Subcloning (.pdf, 488kb) Introduction T-Vector Systems Giving Blunt-Ended DNA an A-Tail for T-Vector Subcloning Subcloning with RE Sites Subcloning using PCR Primers Containing Restriction Sites Ordering Information 35 Chapter 3: Transforming Bacteria (.pdf, 366kb) Properties of E. coli Strains for Subcloning Ready-to-Use Competent Cells Determining Transformation Efficiency of Competent Cells Transforming Ligation Reactions Media and Solutions 43 Chapter 4: Screening for Recombinants (.pdf, 431kb) Introduction Colony PCR Go Directly to Gel Screening by Plasmid Minipreps and RE Digests Plasmid Minipreps Troubleshooting Subcloning Experiments Ordering Information 49 Chapter 5: Technical Appendix (.pdf, 274kb) Restriction Enzyme Activity in 10X Buffers, Reaction Temperature and Heat Inactivation Isoschizomers Compatible Ends Site-Specific Methylation Sensitivity of Promega Restriction Enzymes Restriction Enzyme Buffer Composition Copy Number of Commonly Used Plasmids Star Activity Genotypes of Frequently Used Bacterial Strains Genetic Markers in E. coli Nucleic Acid Calculations Formulas for DNA Molar Conversions http://www.promega.com/guides/su ... Subcloning_ntbk.pdf 三、Beginning Molecular Biology Laboratory Gudie 新手入门的好教材，非常全面，非常亲切。值得收藏。 http://www.research.umbc.edu/~jwolf/method1.html 内容介绍： CHAPTER 1: General Laboratory Methods Safety Procedures Preparation of Solutions Disposal of Buffers and Chemicals Equipment Micropipets Using a pH meter Autoclave operating procedures Operating instructions for spectrophotometer Working with DNA Sterile Technique CHAPTER 2: Instructions for Notebook Keeping CHAPTER 3: Computer User's Guide UNIX commands File Transfer The World Wide Web The Wisconsin Package - gcg. CHAPTER 4: Molecular Biology Methods M.1: Preparation of genomic DNA from bacteria M.2: PCR amplification of DNA M.3: Restriction enzyme digestion of DNA M.4: Phenol/chloroform extraction of DNA M.5: Ethanol precipitation of DNA M.6: Agarose gel electrophoresis M.7: Transformation of E. coli by electroporation M.8: Wizard PCR preps DNA purification system M.9: Alternate method for purifying DNA from agarose gels M.10: Transfection of mammalian cells using Lipofectamine (LTI) M.11: Southern blotting M.12: RT-PCR Protocol M.13: Preparation of sequencing gels M.14: Isolation of RNA from mammalian cells using RNAZOL (Teltest) CHAPTER 5: Tissue Culture Methods Types of cells grown in culture Work area and equipment Preservation and storage Maintenance Safety considerations Tissue culture methods Determining cell counts 四、Cloning Enzymes Cloning Enzymes, one in the Enzyme Resource Guide series, highlights those enzymes important in nucleic acid cloning procedures. Enzymes that modify nucleic acids provide the foundation for many molecular biology techniques: enzymes that are used to synthesize, degrade, join or remove portions of nucleic acids in a controlled and generally defined manner. Specific features of the in vivo functions of these enzymes have been exploited in vitro to provide many of the protocols currently used in nucleic acid manipulations.This guide mainly is targeted Ligases, Kinases and Phosphatases. Table of Contents Page Enzyme Activities Diagram The Cloning Enzymes Gene Cloning Ligases Kinases and Phosphatases RecA Protein and AgarACE® Enzyme Technical Appendix http://www.promega.com/guides/cloning_guide/cloningenz.pdf 五、Nucleic Acid Purification Systems The isolation and purification of DNA is crucial to many applications in molecular biology and techonogy. Over the past decade, the purification of DNA has evolved from a multi-step process involving organic chemicals to a much simpler process, that is safer and faster. Promega continues to develop new DNA purification products to meet the diverse and changing needs of today's life science researcher. This Nucleic Acid Purification Systems guide presents our many new and innovative products for use in DNA and RNA purification. Table of Contents Page Genomic DNA Plasmid DNA Fragment DNA Total RNA http://www.promega.com/guides/nadp_guide/napd_guide.pdf 六、 RNA Analysis Notebook RNA analysis is the starting point for many molecular biology procedures. The RNA Analysis Notebook provides an introduction to RNA procedures such as purifying RNA, amplifying via RT-PCR, making RNA in vitro and analyzing RNA by microarray. In addition, the RNA Analysis Notebook highlights products for performing these procedures. Table of Contents Working with RNA Purifying RNA and mRNA Amplifying RNA with RT-PCR Analyzing RNA with Microarrays Making RNA in vitro Silencing RNA in vivo (RNAi) http://www.promega.com/guides/rna_guide/rna_gde.pdf 七、DNA Analysis Notebook DNA analysis is the starting point for many molecular biology procedures. The DNA Analysis Notebook provides an introduction to DNA procedures such as purifying genomic DNA, amplifying via PCR, retrieving DNA from PCR and cloning PCR DNA. In addition, the DNA Analysis Notebook features Promega products for performing these procedures. Contents Table of Contents Genomic DNA Purification Amplifying DNA PCR Clean-Up Cloning PCR DNA DNA Analysis Tools http://www.promega.com/guides/dna_guide/dna_guide.pdf |
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