PCR-induced (ligase-free) subcloning: a rapid reliable method to subclone polymerase chain reaction (PCR) products

Polymerase Chain Reaction (PCR)

The polymerase chain reaction (PCR) is a powerful and widely used technique that has greatly advanced our ability to analyze genes. Genomic deoxyribonucleic acid (DNA) present in cells contains many thousands of genes. This makes it difficult to isolate and analyze any individual gene. PCR allows specific DNA sequences, usually corresponding to genes or parts of genes, to be copied from genomic...

Polymerase Chain Reaction (PCR): A Short Review

was awarded the Nobel prize in Chemistry along with Michael Smith for his work on PCR2. The PCR is commonly carried out in a reaction volume of 10-200 μl in small reaction tubes (0.2-0.5 ml volumes) in a thermal cycler. The thermal cycler heats and cools the reaction tubes to achieve the temperatures required at each step of the reaction. Many modern thermal cyclers make use of the Peltier effe...

Simplified method for ligase-free cloning of PCR products.

Cloning of polymerase chain reaction (PCR) products into plasmid vectors by conventional methods, relying either on recognition sequences built into the primers or taking advantage of the template-independent terminal transferase activity of Taq DNA polymerase to add an extra adenine to the 3′ end of the product, are often difficult (1,10,12). In recent years, numerous cloning strategies (10,13...

Optimized conditions for cloning PCR products into an XcmI T-vector.

products. BioTechniques 12:28-30. 10.Scharf, S.J., G.T. Horn and H.A. Erlich. 1986. Direct cloning and sequence analysis of enzymatically amplified genomic sequences. Science 233:1076-1078. 11.Shuldiner, A.R., L.A. Scott and J. Roth. 1990. PCR-induced (ligase free) subcloning: a rapid reliable method to subclone polymerase chain reaction (PCR) product. Nucleic Acids Res. 18:1920. 12.Starr, L. a...

Rapid and reliable cloning of PCR products.