Increase of cloning efficiencies by using high-performance liquid chromatography-purified vectors and linkers.
High-performance liquid chromatography (HPLC) was applied to the purification of vector and linker DNAs used in molecular cloning experiments. The data clearly demonstrate that HPLC (Blue Column from LKB/TSK G 4000 SW) based on steric exclusion is a very rapid and convenient method for the size separation of DNA fragments that are virtually free of any contamination. Application of vector and linker DNAs, purified by this method, enabled us to clone nanogram amounts of avian myeloblastosis virus RNA, which was used for the evaluation of a recently described cloning procedure.
D000818:Animals; D009189:Avian Myeloblastosis Virus / Q000378:metabolism; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D009690:Nucleic Acid Conformation; D010957:Plasmids; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000096:biosynthesis; D012367:RNA, Viral / Q000302:isolation & purification*
D004274:DNA, Recombinant; D012333:RNA, Messenger; D012367:RNA, Viral; D012313:RNA