PCR-Mediated Mutagenesis in Sequences Recalcitrant to Homogeneous Amplification

Random mutagenesis by whole-plasmid PCR amplification.

Random mutagenesis has become a powerful means of studying the effects of a large number of permutations of a particular DNA sequence. As a prime example, libraries of randomized cDNA clones, when translated into their corresponding proteins, can be useful in investigating the functional contributions of a mutagenized region to the overall properties of a protein. Existing molecular cloning tec...

Recombinant DNA sequences generated by PCR amplification.

The polymerase chain reaction (PCR) has greatly enhanced the field of molecular biology by making numerous regions of the genome (coding and noncoding), in both extant and extinct taxa, accessible for detailed analysis. PCR is especially well suited for applications in systematic biology because conserved regions that flank variable portions of the genome can be used as primer sites for the amp...

Versatile PCR-mediated insertion or deletion mutagenesis.

Localized in situ amplification (LISA). PCR amplification of nucleic acid sequences in tissue sections.

The polymerase chain reaction (PCR) has revolutionized the manner in which molecular biologists are able to examine nucleic acids by offering an extremely sensitive mechanism for amplifying specific target sequences. The combination of increased sensitivity owing to the amplification process and increased specificity owing to primer sequences with subsequent analyses by DNA sequencing, restrict...

Betaine improves the PCR amplification of GC-rich DNA sequences.

Betaine improves the co-amplification of the two alternatively spliced variants of the prostate-specific membrane antigen mRNA as well as the amplification of the coding cDNA region of c-jun. It is suggested that betaine improves the amplification of these genes by reducing the formation of secondary structure caused by GC-rich regions and, therefore, may be generally applicable to ameliorate t...