A Laboratory Guide to Genomic Sequencing

Inhaltsverzeichnis

• I. Introduction.
• A Safety Considerations.
• B Introduction.
• C The Principle of Genomic Sequencing.
• II Theoretical Background.
• A Basic Theoiy of Genomic Sequencing.
• B Flow Diagram.
• III Experimental.
• 1 Isolation of Genomic DNA.
• 2 Restriction Digest of Genomic DNA.
• 3 Chemical Sequencing Reactions on Restricted DNA.
• 4 Separation of Reaction Products on a Sequencing Gel.
• 5 Electrotransfer to Nylon Membranes.
• 6 Immobilization of DNA on a Nylon Membrane.
• 7 Prehybridization and Hybridization of Immobilized DNA with Labeled Single-Stranded DNA Probes.
• 8 Processing of the Hybridized Filters.
• 9 Autoradiography and Photography.
• 10 Cloning of DNA Probe in M13.
• 11 Large-Scale Preparation of Cloned DNA in M13.
• 12 Synthesis of Oligonucleotide Primers and Single-Stranded Labeled Probes.
• 13 Purification of Labeled Single-Stranded Probes.
• IV Trouble-Shooting Guide and Examples.
• V Appendix.
• VI Bibliography.