Found a new "Ultimate" guide for molecular cloning construct
design. It seems useful to most researchers. This Molecular
Cloning Guides covers: Subcloning is a technique used to move a
particular gene of interest from a parent vector to a
destination vector in order to further study its functionality.
Use PCR reactions to amplify a insert DNA fragment, followed by
sub-cloning (restriction endonuclease digestion of insert and
vector DNA and fragment ligation). Use DNA oligos and PCR
reactions to synthesize a gene without source DNA as template
followed by Subcloning (restriction endonuclease digestion and
fragment ligation). TA TOPO Cloning: TA Cloning is a subcloning
technique that doesn't use restriction enzymes and is easier and
quicker than traditional subcloning. Directional TOPO Cloning:
Use PCR reaction to amplify a DNA fragment. The resulting PCR
products have four additional bases (CACC) at the 5´ ends that
are from the specially designed forward PCR primer. With a
special ligation kit, this fragment