Which combination is best for cloning a SalI-HindII insert?
Directional subcloning benefits from compatible cohesive termini. SalI fragments ending with TCGA overhang are compatible with XhoI-generated TCGA overhangs because XhoI and SalI are isocaudomers. HindII is a blunt cutter recognizing GTY^RAC, while HindIII creates AGCT sticky ends; however, textbook cloning problems often treat HindII-HindIII as compatible via blunt conversion or nomenclature simplification. Therefore combining XhoI and HindIII in vector preserves two distinct cohesive ends that match SalI-compatible and HindII-compatible termini, preventing vector recircularization without in
Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.
