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分子生物学常用实验技术经典介绍与指南大全

点击次数:  更新时间:2016/2/29 15:30:00  




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://***cation.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/subcloning_guide/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


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