روشي ايده ال براي توليد حامل TA مناسبترين ناقل براي كلون كردن محصول PCR

A choice procedure for production of TA vector, the best vehicle for cloning of PCR product

PCR is an in vitro technique which allows the amplification of a specific DNA region that lies between two regions of a known DNA sequence. PCR product must be cloned to provide a permanent source of the amplified DNA fragment. One of the necessary tools for cloning, is vector. PUC19 is a vector that is widely used in cloning procedures. In this study, for production of TA vector, pUC19 was digested with Smal restriction enzyme. A single T nucleotide was added to the 3ʹ ends of blunt-cut pUC19 using dTTP and Taq DNA polymerase, which had terminal transferase activity. In this way, TA vector was created. The use of Taq DNA polymerase leads to add a single A residue at the 3ʹ ends of the PCR product. The T-A overhangs in vector/ PCR product, respectively, facilitate the ligation reaction and cloning of PCR product. So the TA vector produced by this method, can be efficiently used for cloning of PCR product.