Basic molecular biology : essential techniques /
| Main Author: | |
|---|---|
| Format: | Book |
| Language: | English |
| Published: |
Chichester ; New York :
Wiley,
[1999]
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| Series: | Essential techniques series.
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| Subjects: |
Table of Contents:
- DNA structure
- Denaturation and renaturation (hybridization) of
- DNA
- RNA structure
- Enzymes used in molecular biology
- Restriction endonucleases
- DNA polymerases
- RNA polymerases
- Nucleases
- End-modification enzymes
- Ligases
- DNA cloning
- Setting up a molecular biology laboratory
- Preparation of DNA
- Methods available
- Protocols
- Preparation of cell pellet and cell lysis
- Phenol extraction of DNA
- Ethanol precipitation of DNA
- Removal of RNA by ribonuclease treatment
- Final purification steps
- Assessment of DNA concentration and DNA quality
- Preparation of RNA
- Methods available
- Protocols
- Preparation of cell pellet
- Preparation of cytoplasmic RNA
- Preparation of whole cell RNA by ultracentrifugation method
- Preparation of whole cell RNA by Chomczynski-Sacchi method
- Assessment of RNA concentration and RNA quality
- Selection of poly(A)* RNA
- Preparation of plasmid DNA
- Methods available
- Protocols
- Transformation of bacteria with plasmid
- Growth of bacteria and amplification of plasmid
- Small-scale preparation of plasmid DNA
- Large-scale preparation of plasmid DNA
- Isolation of cloned gene sequences
- DNA analysis by restriction enzyme digestion and Southern blotting
- Methods available
- Protocols
- Restriction endonuclease digestion
- Concentration of DNA by ethanol precipitation
- Agarose gel electrophoresis
- Southern blotting
- Radiolabeling of DNA probe
- Hybridization of blot
- Washing of blot
- 5'-end radiolabeling of DNA molecular weight markers
- DNA analysis by PCR
- Methods available
- Protocols
- PCR reaction
- Analysis of PCR products by agarose gel electrophoresis
- RNA analysis by Northern blotting
- Methods available
- Protocols
- Formaldehyde-agarose gel electrophoresis
- Northern blotting and hybridization to radiolabelled DNA probe
- RNA analysis by RNase protection
- Methods available
- Protocols
- Preparation of riboprobe
- RNase protection assay
- Analysis of products by polyacrylamide gel electrophoresis
- RNA analysis by RT-PCR and differential display
- Methods available
- Protocols
- RT-PCR reaction and analysis of products
- Differential display
- Analysis of protein-DNA interactions by gel shift assay
- Methods available
- Protocols
- Preparation of protein extract
- Gel shift assay
- Gene transfection
- Methods used
- Protocols
- Transient transfection
- Stable transfection
- Appendix A: recipes for routine solutions
- Appendix B: Sources of reagents and equipment.