Basic Cloning Procedures
Springer Berlin Heidelberg, Mar 10, 1998 - Science - 169 pages
This manual introduces the reader to basic methods used in the isolation, cloning and analysis of genetic material. The protocols include RT-PCR amplification, gene cloning, hybridization analysis and sequencing of nucleic acids, PCR-based site-specific mutagenesis, analysis of protein DNA-specific interaction, cell-free protein synthesis and product electrophoretic and immunological analysis. Each protocol includes short background information, a detailed description of the necessary materials, step-by-step procedures, a troubleshooting guide and useful practical hints.
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acetate acrylamide agarose gel Amersham amplification analysis aqueous phase autoradiography bands binding bromophenol blue cells Centrifuge cloning coli concentration containing denaturation digestion DNA bending DNA fragments DNA polymerase DNA sequencing DNA template DNase dNTP double-stranded EDTA efficiency EMSA ethanol ethidium bromide Fermentas filter footprinting gel electrophoresis gene genomic DNA hybridization solution Incubate lanes loading membrane method microcentrifuge tube molecular molecules mRNA mutagenesis mutant primer NaCl Northern blotting Note nuclease nucleic acids nucleotide oligonucleotide overnight PAAG PCR product pellet Pharmacia plasmid plate poly(A)+mRNA Pra/A preparation probe Procedure Promega promoter protein protein-DNA complexes protocols purification radioactive RafR reaction mixture repressor restriction enzymes Resuspend reticulocyte ribonuclease RNase room temperature sample staining sterile supernatant synthesis T7 RNA polymerase template DNA tion total RNA transcription transfer Tris-HCl vector vitro translation volume vortexing wash wheat germ extract X-ray film xylene cyanol