1. Other Molecular Techniques

2. Blotting

8. Probe synthesis Nick Translation –DNA template preparation –Nick template with DNase I –Fill in gaps with DNA polymerase and labeled nucleotides –Denature and hybridize Random Priming –DNA template preparation –Anneal with random hexamers –Primer extend with DNA polymerase and labeled nucleotides –Denature and hybridize

9. Probe synthesis End labeling –Prepare template –End label with labeled ATP and polynucleotide kinase –Denature and hybridize RNA probes –Clone template into a T7, T3 or Sp6 vector –Restriction cut to linearize –RNA polymerization with labeled rNTPs –Denature and hybridize

12. Dot Blotting

13. Dot Blot Scheme Isolate RNA Make dilution series Heat and dot onto membranes Make labeled probe Hybridize Wash Autoradiograph/detect

18. Transcription Mapping

24. Promoter Characterization

36. In vitro mutagenesis

40. Protein Characteristics

43. Protein Structure 1º- Primary –Linear sequence of amino acids/peptide bonds 2º- Secondary –Hydrogen bonds -  helix &  sheet 3º- Tertiary- organized into subunit domains –Hydrogen, ionic & disulfide bonds 4º- Quaternary- organization of multiple domains into a multi-subunit protein –Hydrogen, ionic,metallo & disulfide bonds

58. Physical Separation Techniques

59. Centrifugation

71. Equilibrium Centrifugation Ultra Centrifuges CsCl gradients that self-form Separates on the basis of buoyant density Molecules migrate to a specific buoyant density at equilibrium DNA migrates differentially depending on AT/GC composition RNA may pellet

72. Column Chromatography

80. Protein Electrophoresis

87. SDS Gel Electrophoresis

89. Isoelectric Focusing/ 2 Dimensional Gels

95. Western and Electroblotting

100. Peptide Mapping

105. Isotope Labeling

108. X Ray Crystallography