a highly efficient cloning vector was constructed for cloning pcr products by inserting an 80 bp dna fragment into pgem-5zf (+) vector. the xcm i digestion of this vector gave rise to a 3’ overhanging deoxythymidine offering the possibility of cloning pcr products with 3' adenosine overhang created by taq dna polymerase. furthermore, two ecor i sites were added to the construct for identificati...

A variety of methods have been developed for cloning PCR products, including blunt-end cloning (1), restriction cut back (2), ligation-independent cloning (3), uracil DNA–glycosylase (UDG) treatment of uracil-containing deoxyoligonucleotide primers (4,5) and TA cloning (6–8). Blunt-end cloning of PCR products often requires treatment of PCR products to polish the ends (9). Even with treatments,...

knowing the nucleotide sequence of the cholera toxin operon, we designed oligonucleotide primers for its-pcr amplification from local clinical isolates of v. cholerae. the resulting amplification product was cloned in a common puc18 vector. subsequently, a part of this operon encoding the cholera toxin bsubunit (ctb) was reamplified and cloned between the bamh1 and ecor1 sites of the same vecto...

A rapid DNA cloning system is a research interest of many scientists. TA cloning is one of the methods used for the cloning of PCR-amplified DNA molecules. The TA cloning method is a convenient and labor-saving replacement to traditional, restriction enzyme-mediated cloning strategies. A T-vector called pBlueskript ΙΙ SK-1 with the lethal gene ccdB was designed to construct a positive selection...

............................................................................................................................. 3 INTRODUCTION.................................................................................................................... 3 AIM..........................................................................................................................................