Molecular biology

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Revision as of 08:32, 21 July 2009

Reagents & Protocols

recipes

1. Chemicals
2. Commonly used reagents and equipment in molecular biology (PDF)
3. Concentration of antibiotics commonly used
4. For DNA extraction (to remove some protein contaminants & inactivate enzymes): Phenol:Chloroform=1:1(v:v) solution
5. DNA sequencing check list
   1. quality score
   2. good sequencing result vs. bad sequencing result (see below example)
      

      

protocols

• PCR - General rules for primer design
• Genomic DNA extraction
• Competent cell preparation BSGC protocol (PDF)
• Electrocompetent cell Protocol (PDF)
• LIC cloning introduction LIC introduction - Ligation Independent Cloning
  • LIC cloning protocol (Berkeley Structural Genomics Center)
    • Vector preparation
    • Insert preparation
    • LIC transformation:
      • mix insert and vector in appropriate amounts
      • incubate 5~30 min(20 min) at room temperature
      • do transformation in E. coli (recommend to use electro-competent cells)
• NCBE practical protocols: very good illustration for undergraduate students
• Conjugation
  • Donor: E. coli S17-1

Tools

webtools

1. Oligo Tm calculation

softwares

1. NIH Image: Gel analysis software
2. Serial Cloner: DNA sequence management (PCR, restriction maps, etc.)
3. BioX (Mac OS X only)
4. Biological sequence editor Bioedit
5. Jmol