Molecular biology
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===protocols=== | ===protocols=== | ||
#[[PCR primer|PCR]] - General rules for primer design | #[[PCR primer|PCR]] - General rules for primer design | ||
- | ##[[media:takarPCRprotocol.pdf|Takara PCR protocol] | + | ##[[media:takarPCRprotocol.pdf|Takara PCR protocol]] |
#[[Genomic DNA extraction]] | #[[Genomic DNA extraction]] | ||
#Competent cell preparation [[media:competentcell.pdf|BSGC protocol (PDF)]] | #Competent cell preparation [[media:competentcell.pdf|BSGC protocol (PDF)]] |
Revision as of 08:10, 14 August 2008
Contents |
Reagents & Protocols
recipes
- 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)
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)
Tools
Webtools
Softwares
- NIH Image: Gel analysis software
- Serial Cloner: DNA sequence management (PCR, restriction maps, etc.)
- BioX (Mac OS X only)
- Biological sequence editor Bioedit
- Jmol test