Library Science

This volume provides a set of protocols for the analysis of cloned genes. A set of techniques is described for the basic manipulation and mutagenesis of the cloned sequences. The rest of the volume is organized into a collection of techniques appropriate to the analytical route to be investigated; these include genome structure, sequence variation, gene expression, and protein function. Finally, a number of methods are described to take the step from the first gene to those with which it interacts.

Authors: Harwood, Adrian J.

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Table of Contents

1. Transformation of Bacteria by Electroporation (pp. 1-8)
   Drury, Lucy
2. Direct Cloning of (lambda)gt11 cDNA Inserts Into a Plasmid Vector (pp. 9-18)
   Poulin, Matthew L.; Chiu, Ing-Ming
3. PCR Cloning Using T-Vectors (pp. 19-34)
   Trower, Michael K.; Elgar, Greg S.
4. Thermal Cycle Dideoxy DNA Sequencing (pp. 35-46)
   Slatko, Barton E.
5. Ordered Deletions Using Exonuclease III (pp. 47-56)
   Clark, Denise; Henikoff, Steven
6. Site-Directed Mutagenesis Using a Double-Stranded DNA Template (pp. 57-66)
   Viville, Stephane
7. Site-Directed Mutagenesis Using a Uracil-Containing Phagemid Template (pp. 67-78)
   Trower, Michael K
8. Construction of Linker-Scanning Mutations by Oligonucleotide Ligation (pp. 79-86)
   Hobson, Grace M.; Harlow, Patricia P.; Benfield, Pamela A.
9. Construction of Linker-Scanning Mutations Using the Polymerase Chain Reaction (pp. 87-96)
   Harlow, Patricia P.; Hobson, Grace M.; Benfield, Pamela G
10. Localized Random Polymerase Chain Reaction Mutagenesis (pp. 97-112)
    Lerner, Claude G.; Inouye, Masayori
11. The Simultaneous Isolation of RNA and DNA from Tissues and Cultured Cells (pp. 113-120)
    Merante, Frank; Raha, Sandeep; Reed, Juta K; Proteau, Gerald
12. Physical Mapping of the Human Genome by Pulsed Field Gel Electrophoresis (pp. 121-134)
    Boultwood, Jacqueline
13. Field Inversion Gel Electrophoresis (pp. 135-146)
    Heller, Christoph
14. Enhanced Chemiluminescent Detection of Horseradish Peroxidase Labeled Probes (pp. 147-162)
    Durrrant, Ian
15. Nonradioactive Oligonucleotide Probe Labeling (pp. 163-176)
    Durrant, Ian; Fowler, Sue
16. Analysis of DNA Restriction Enzyme Digests by Two-Dimensional Electrophoresis in Agarose Gels (pp. 177-186)
    Kuff, Edward L.; Mietz, Judy A.
17. Inverse Polymerase Chain Reaction (pp. 187-196)
    Hartl, Daniel L.; Ochman, Howard
18. Use of Silver Staining to Detect Nucleic Acids (pp. 197-204)
    Mitchell, Lloyd G.; Bodenteich, Angelika; Merril, Carl R
19. A Nonradioactive Method for the Detection of Single-Strand Conformational Polymorphisms (SSCP) (pp. 205-210)
    Ainsworth, Peter J.; Rodenhiser, David I.
20. Temperature Gradient Gel Electrophoresis (TGGE) for the Detection of Polymorphic DNA and RNA (pp. 211-228)
    Henco, Karsten; Harders, Jutta; Wiese, Ulrich; Riesner, Detlev
21. TGGE in Quantitative PCR of DNA and RNA (pp. 229-236)
    Kang, Jie; Harders, Jutta; Riesner, Detlev; Henco, Karsten
22. The PGK-PCR Clonality Assay (PPCA) (pp. 237-246)
    Busque, Lambert; Gilliland, D. Gary
23. Direct Sequencing of PCR Products (pp. 247-256)
    Phear, Geraldine A.; Harwood, Janet
24. The Use of Riboprobes for the Analysis of Gene Expression (pp. 257-272)
    Belin, Dominique
25. Quantification of Absolute Amounts of Cellular Messenger RNA by RNA-Excess Solution Hybridization (pp. 273-280)
    Srivastava, Rai Ajit K.; Schonfeld, Gustav
26. Measurements of Rate of Transcription in Isolated Nuclei by Nuclear ??°Run-Off??² Assay (pp. 281-288)
    Srivastava, Rai Ajit K.; Schonfeld, Gustav
27. An RNA Polymerase II In Vitro Transcription System (pp. 289-298)
    Mantovani, Roberto
28. S1 Mapping Using Single-Stranded DNA Probes (pp. 299-306)
    Viville, Stephane; Mantovani, Roberto
29. Single Primer-Mediated Polymerase Chain Reaction (pp. 307-316)
    Myers, Rene L.; Chiu, Ing-Ming
30. In Vivo DNA Footprinting by Linear Amplification (pp. 317-330)
    Saluz, Hans Peter; Jost, Jean-Pierre
31. DNA photofootprinting with Rh(phi)2bpy3+ (pp. 331-338)
    Klakamp, Scott L.; Barton, Jacqueline K
32. The Gel Retardation Assay (pp. 339-348)
    Scott, Valerie; Clark, Andrew R; Docherty, Kevin
33. The Southwestern Assay (pp. 349-362)
    Philippe, Jacques
34. Cloning DNA Binding Proteins from cDNA Expression Libraries Using Oligonucleotide Binding Site Probes (pp. 363-370)
    Cowell, Ian G.; Hurst, Helen C.
35. 6xHis-Ni-NTA Chromatography as a Superior Technique in Recombinant Protein Expressiod/Purification (pp. 371-388)
    Crowe, Joanne; Dobeli, Heinz; Gentz, Reiner; Hochuli, Erich; Stuber, Dietrich; Henco, Karsten
36. Production of 35S-Labeled Proteins in E. coli and Their Use as Molecular Probes (pp. 389-396)
    Lydan, Michael A.; O???Day, Danton H.
37. Preparation and Ligand Screening of a (lambda)gt11 lysogen Library (pp. 397-408)
    Mutzel, Rupert

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