Molecular biology

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Revision as of 05:10, 28 July 2009

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

good vs. bad sequencing example

PCR - General rules for primer design
Genomic DNA extraction
Competent cell preparation BSGC protocol (PDF)
- Easy & rapid competent cell preparation reference link - by HJKo
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

@@ Line 14: / Line 14: @@
 *[[Genomic DNA extraction]]
 *Competent cell preparation [[media:competentcell.pdf|BSGC protocol (PDF)]]
-*Easy & rapid competent cell preparation [http://www.personal.psu.edu/faculty/d/s/dsg11/labmanual/DNA_manipulations/Comp_bact_by_TSS.htm reference link] - by HJKo
+**Easy & rapid competent cell preparation [http://www.personal.psu.edu/faculty/d/s/dsg11/labmanual/DNA_manipulations/Comp_bact_by_TSS.htm reference link] - by HJKo
 *Electrocompetent cell [[media:ElectrocompetentEcoli.pdf|Protocol (PDF)]]
 *LIC cloning introduction [[media:LICnovagen.pdf|LIC introduction]] - Ligation Independent Cloning