Cloning protocols/tips by Astrid. PCR and primer design, Extracting DNA from gel, Restriction of vector, Ligation, and Transformation. CalTech - [Read Cloning protocols/tips by Astrid PDF]
Agarose gel purification,
Annealing and extending, oligonucleotides, Ethanol Precipitation, Ligation
Miniprep, Oligonucleotide purification, Recovering DNA bands, Restriction digest
Gene Clean. The Hu Lab. - [Read DNA cloning Procedures]
Preparation of oligo solutions
PCR experiments, Digestion of insert DNA, Digestion and dephosphorylation of vector DNA, Ligation of DNA fragments with sticky ends, Ligation of DNA fragments with blunt ends,
Preparation of chemically competent E. coli - [Read EMBL Cloning protocols]
The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.