Molecular biology
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**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)]] | *Electrocompetent cell [[media:ElectrocompetentEcoli.pdf|Protocol (PDF)]] | ||
| - | + | ====LIC cloning==== | |
| + | *Reference: | ||
<biblio>LIC pmid=2235490</biblio> | <biblio>LIC pmid=2235490</biblio> | ||
| + | *LIC cloning introduction [[media:LICnovagen.pdf|LIC introduction]] - Ligation Independent Cloning | ||
**LIC cloning protocol (Berkeley Structural Genomics Center) | **LIC cloning protocol (Berkeley Structural Genomics Center) | ||
***[http://www.strgen.org/protocols/lic-vector-prep.pdf Vector preparation] | ***[http://www.strgen.org/protocols/lic-vector-prep.pdf Vector preparation] | ||
Revision as of 13:12, 23 November 2010
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Reagents & Protocols
recipes
- 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)
protocols
- 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
- Reference:
Error fetching PMID 2235490:
- Error fetching PMID 2235490:
- 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)
- LIC cloning protocol (Berkeley Structural Genomics Center)
- NCBE practical protocols: very good illustration for undergraduate students
- Conjugation
- Donor: E. coli S17-1
- E. coli knockout by the Wanner method (Lambda Red Recombinase)
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
