USER fusion: a rapid and efficient method for simultaneous fusion and cloning of multiple PCR products

USER fusion: a rapid and efficient method for simultaneous fusion and cloning of multiple PCR products

We present a method that allows simultaneous fusion and cloning of multiple PCR products in a rapid and efficient manner. The procedure is based on the use of PCR primers that contain a single deoxyuridine residue near their 5' end. Treatment of the PCR products with a commercial deoxyuridine-excision reagent generates long 3' overhangs designed to specifically complement each other. The combin...

Hetero-stagger cloning: efficient and rapid cloning of PCR products.

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,...

Inverse Fusion PCR Cloning

Inverse fusion PCR cloning (IFPC) is an easy, PCR based three-step cloning method that allows the seamless and directional insertion of PCR products into virtually all plasmids, this with a free choice of the insertion site. The PCR-derived inserts contain a vector-complementary 5'-end that allows a fusion with the vector by an overlap extension PCR, and the resulting amplified insert-vector fu...

A REPORT OF RAPID AND EFFICIENT METHOD FOR MITOCHONDRIAL DNA

Based on the similarity of prokaryotic DNA to that of mitochondrial DNA, a rapid and efficient method for DNA purification from mitochondrial extracts obtained from different sources, was carried out based on the modification method described by Marmur (1983) which resulted in a high degree of purity (A260/A280=2). In addition we have applied the same technique for the first time to purify...

Rapid and reliable cloning of PCR products.