PCR amplification of megabase DNA with tagged random primers (T-PCR)

PCR amplification of megabase DNA with tagged random primers (T-PCR).

A very sensitive and specific method for the random amplification of whole DNA molecules and genomes ranging from 400 base pairs (bp) to 40 Megabase (Mb) is described. This simple, two step PCR (1-3) strategy utilizes tagged random primers that consist of a pool of all possible 3' sequences for binding to the target DNA and a constant 5' region for the detection of incorporated primers. As litt...

Panero and Crozier: Asteraceae Chloroplast Dna Primers Primers for Pcr Amplification of Asteraceae

Primers for the amplification and sequencing of DNA fragments from chloroplast genes and non-coding regions are provided to facilitate molecular phylogenetic studies aimed at building a tree-of-life for the Asteraceae. The primers reported here have been extensively tested and some empirical guidelines are included to facilitate their use.

PCR with arbitrary primers

Emulsion PCR: A High Efficient Way of PCR Amplification of Random DNA Libraries in Aptamer Selection

Aptamers are short RNA or DNA oligonucleotides which can bind with different targets. Typically, they are selected from a large number of random DNA sequence libraries. The main strategy to obtain aptamers is systematic evolution of ligands by exponential enrichment (SELEX). Low efficiency is one of the limitations for conventional PCR amplification of random DNA sequence library in aptamer sel...

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...