Library Science

An essential core collection of the latest molecular and genetic techniques for cloning, subcloning, sequencing, PCR, protein expression, and much more. Each protocol represents a time-tested, step-by-step recipe that creates an understanding of the procedure, easily reproducible results, and confidence that the procedure will work. The collection includes not only many updated and improved classic techniques, but also a powerful group of advanced methods that point to future progress, among them nonisotopic DNA labeling, silver staining, and automatic sequencing. This excellent bench companion will help those who need to learn for the first time how to conduct research on the molecular biology of nucleic acids or those who need to broaden their competence and laboratory skills. Even highly skilled researchers will find many time-saving techniques.

basic-dna-and-rna-protocols.pdf

Table of Contents

1. The Simultaneous Isolation of RNA and DNA from Tissues and Cultured Cells (pp. 3-10)
   Merante, Frank; Raha, Sandeep; Reed, Juta K.; Proteau, Gerald
2. Restriction Endonuclease Digestion of DNA (pp. 11-16)
   Smith, Duncan R.
3. Agarose Gel Electrophoresis (pp. 17-22)
   Smith, Duncan R.
4. Capillary Blotting of Agarose Gels (pp. 23-26)
   Smith, Duncan R.; Murphy, David
5. Random Primed 32P-Labeling of DNA (pp. 27-30)
   Smith, Duncan R.
6. Hybridization and Competition Hybridization of Southern Blots (pp. 31-40)
   Kelsell, Rosemary E.
7. Utilization of DNA Probes with Digoxigenin-Modified Nucleotides in Southern Hybridizations (pp. 41-52)
   Helentjaris, Tim; McCreery, Tom
8. The Preparation of Fluorescein-Labeled Nucleic Acid Probes and Their Detection Using Alkaline Phosphatase-Catalyzed Dioxetane Chemiluminescence (pp. 53-64)
   Cunningham, Martin; Harris, Martin
9. Monitoring Incorporation of Fluorescein into Nucleic Acid Probes Using a Rapid Labeling Assay (pp. 65-66)
   Cunningham, Martin
10. The Preparation of Horseradish Peroxidase-Labeled Nucleic Acid Probes and Their Detection Using Enhanced Chemiluminescence (pp. 67-76)
    Harvey, Bronwen; Durrant, Ian; Cunningham, Martin
11. 3’-End Labeling of Oligonucleotides with Fluorescein-11-dUTP and Enhanced Chemiluminescent Detection (pp. 77-82)
    Cunningham, Martin; Durrant, Ian; Harvey, Bronwen
12. The Preparation of Riboprobes (pp. 83-92)
    Belin, Dominique
13. Native Polyacrylamide Gel Electrophoresis (pp. 93-96)
    Harwood, Adrian J.
14. Use of Silver Staining to Detect Nucleic Acids (pp. 97-104)
    Mitchell, Lloyd G.; Bodenteich, Angelika; Merril, Carl R.
15. End-Labeling of DNA Fragments (pp. 105-110)
    Harwood, Adrian J.
16. Northern Blot Analysis (pp. 113-128)
    Krumlauf, Robb
17. RNA Slot Blotting (pp. 129-130)
    Murphy, David
18. The RNase Protection Assay (pp. 131-136)
    Belin, Dominique
19. Primer Extension Analysis of mRNA (pp. 137-146)
    Leonard, Mark W.; Patient, Roger K.
20. S1 Mapping Using Single-Stranded DNA Probes (pp. 147-154)
    Viville, Stephane; Mantovani, Roberto
21. Nonradioactive In Situ Hybridization for Cells and Tissues (pp. 155-167)
    Durrant, Ian
22. In Vitro Packaging of DNA (pp. 171-176)
    Dale, Jeremy W.; Greenaway, Peter J.
23. Construction of Mammalian Genomic Libraries Using lambda Replacement Vectors (pp. 177-190)
    Bateson, Alan N.; Pollard, Jeffrey W.
24. The Production of Double-Stranded Complementary DNA for Use in Making Libraries (pp. 191-198)
    Mayall, Steve; Kirk, Jane
25. Construction of cDNA Libraries (pp. 199-210)
    Burrell, Michael M.
26. Screening lambda Libraries (pp. 211-218)
    Harwood, Janet C.; Harwood, Adrian J.
27. Subcloning Strategies and Protocols (pp. 221-236)
    Taghian, Danielle Gioioso; Nickoloff, Jac A.
28. Purification of DNA Fragments from Agarose Gels Using Glass Beads (pp. 237-240)
    Joly, Etienne
29. Transformation of E. coli (pp. 241-248)
    Tomley, Fiona M.
30. Transformation of Bacteria by Electroporation (pp. 249-256)
    Drury, Lucy
31. Preparation of Plasmid DNA Using Alkaline Lysis (pp. 257-264)
    Joly, Etienne
32. The Rapid Boiling Method for Small-Scale Preparation of Plasmid DNA (pp. 265-268)
    Harwood, Adrian J.
33. Plasmid Preparations with Diatomaceous Earth (pp. 269-272)
    Machesky, Laura
34. Polymerase Chain Reaction (pp. 275-292)
    Delidow, Beverly C.; Lynch, John P.; Peluso, John J.; White, Bruce A.
35. Inverse Polymerase Chain Reaction (pp. 293-302)
    Hartl, Daniel L.; Ochman, Howard
36. Polymerase Chain Reaction with Degenerate Oligonucleotide Primers to Clone Gene Family Members (pp. 303-312)
    Preston, Gregory M.
37. Cloning PCR Products Using T-Vectors (pp. 313-324)
    Dower, Michael K.; Elgar, Greg S.
38. Direct Radioactive Labeling of Polymerase Chain Reaction Products (pp. 325-328)
    McDaniel, Tim; Meltzer, Stephen J.
39. A Rapid PCR-Based Colony Screening Protocol for Cloned Inserts (pp. 329-334)
    Trower, Michael K.
40. Use of Polymerase Chain Reaction to Screen Phage Libraries (pp. 335-339)
    Yu, Lei; Bloem, Laura J.
41. Cloning into M13 Bacteriophage Vectors (pp. 343-348)
    Yu, Qingzhong
42. Ordered Deletions Using Exonuclease III (pp. 349-358)
    Clark, Denise; Henikoff, Steven
43. M13 Phage Growth and Single-Stranded DNA Preparation (pp. 359-362)
    Tomley, Fiona M.
44. Preparation of ssDNA from Phagemid Vectors (pp. 363-366)
    Trower, Michael K.
45. A Rapid Plasmid Purification Method for Dideoxy Sequencing (pp. 367-372)
    Griffin, Annette M.; Griffin, Hugh G.
46. DNA Sequencing Using Sequenase Version 2.0 T7 DNA Polymerase (pp. 373-388)
    Fuller, Carl W.; McArdle, Bernard F.; Griffin, Annette M.; Griffin, Hugh G.
47. Pouring Linear and Buffer-Gradient Sequencing Gels (pp. 389-392)
    Littlebury, Paul
48. Electrophoresis of Sequence Reaction Samples (pp. 393-402)
    Bankier, Alan T.
49. Direct Sequencing of PCR Products (pp. 403-412)
    Harwood, Janet C.; Phear, Geraldine A.
50. Thermal Cycle Dideoxy DNA Sequencing (pp. 413-424)
    Slatko, Barton E.
51. Using the Automated DNA Sequencer (pp. 425-440)
    Du, Zijin; Wilson, Richard K.
52. Terminal Labeling of DNA for Maxam and Gilbert Sequencing (pp. 441-446)
    Pichersky, Eran
53. DNA Sequencing by the Chemical Method (pp. 447-451)
    Pichersky, Eran
54. Site-Directed Mutagenesis of Double-Stranded Plasmids, Domain Substitution, and Marker Rescue by Comutagenesis of Restriction Enzyme Sites (pp. 455-468)
    Nickoloff, Jac A.; Deng, Win-Ping; Miller, Elizabeth M.; Ray, F. Andrew
55. A Protocol for Site-Directed Mutagenesis Employing a Uracil-Containing Phagemid Template (pp. 469-476)
    Trower, Michael K.
56. In Vitro Translation of Messenger RNA in a Rabbit Reticulocyte Lysate Cell-Free System (pp. 477-484)
    Olliver, Louise; Boyd, Charles D.
57. In Vitro Translation of Messenger RNA in a Wheat Germ Extract Cell-Free System (pp. 485-490)
    Olliver, Louise; Grobler-Rabie, Anne; Boyd, Charles D.
58. One-Step Purification of Recombinant Proteins with the 6xHis Tag and Ni-NTA Resin (pp. 491-510)
    Crowe, Joanne; Masone, Brigitte Steude; Ribbe, Joachim

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