Molecular biology

From CSBLwiki

Contents 1 Reagents & Protocols 1.1 recipes 1.2 protocols 2 Softwares

Reagents & Protocols

recipes

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

      

protocols

1. PCR - General rules for primer design
2. Genomic DNA extraction
3. Competent cell preparation BSGC protocol (PDF)
4. Electrocompetent cell Protocol (PDF)
5. LIC cloning introduction LIC introduction - Ligation Independent Cloning

• LIC cloning protocol (Berkeley Structural Genomics Center)

1. Vector preparation
2. Insert preparation
3. LIC transformation:
   1. mix insert and vector in appropriate amounts
   2. incubate 5~30 min(20 min) at room temperature
   3. do transformation in E. coli (recommend to use electro-competent cells)

Softwares

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