PURPOSE This study examined whether retinal pigment epithelial (RPE) cells have the capacity to express transforming growth factor-beta (TGF-beta). Also examined were TGF-beta isoform genes expressed in the RPE cells. METHODS Complementary DNA (cDNA) was generated from polyA+ RNA extracted from human RPE cells in culture, and polymerase chain reaction (PCR) using degenerate and specific prime...

A PCR primer sequence is called degenerate if some of its positions have several possible bases. The degeneracy of the primer is the number of unique sequence combinations it contains. We study the problem of designing a pair of primers with prescribed degeneracy that match a maximum number of given input sequences. Such problems occur when studying a family of genes that is known only in part,...

1Department of Dermatology, Wayne State University, Detroit, Michigan 48201; 2Department of Dermatology, Niigata University, Niigata, Japan As the application of PCR has expanded, more amplified products have been used for cloning and mutagenesis. Although Taq polymerase (from eubacterium Thermus aquaticus) is the most convenient and intensively studied enzyme to date, one of its drawbacks is a...

The multiplex polymerase chain reaction (MP-PCR) is a quick and an inexpensive technique in molecular biology for amplifying multiple DNA loci in a single Polymerase Chain Reaction (PCR). One of the criteria to achieve highly specific reaction products is to keep the concentration of the amplification primers low. In research, the dilemma associated with primer minimization for MP-PCR reactions...

To detect anoxygenic bacteria containing either type 1 or type 2 photosynthetic reaction centers in a single PCR, we designed a degenerate primer set based on the bchY gene. The new primers were validated in silico using the GenBank nucleotide database as well as by PCR on pure strains and environmental DNA.

Amplification of multiple DNA sequences in a single Polymerase Chain Reaction (PCR) experiment, called Multiplex PCR (MPPCR), is a widely known laboratory technique that requires efficient selection of a set of synthetic strings called Degenerate Primers to result in successful PCR products. The Multiple Degenerate Primer Selection Problem (MDPSP) is a fundamental problem in molecular biology a...

Degenerate primers for the [FeFe] hydrogenase (hydA) were developed and used in PCRs to examine hydA in microbial mats that inhabit saltern evaporative ponds in Guerrero Negro (GN), Mexico. A diversity of deduced HydA was discovered that revealed unique variants, which may reflect adaptation to the environmental conditions present in GN.

Eucalyptus species synthesize a wealth of polyketide natural products, but no relevant biosynthetic enzyme has been identified. Degenerate primers designed from conserved regions of fourteen chalcone synthase superfamily enzymes were used to isolate gene fragments from at least five different Type III polyketide synthases (PKSs) in E. camaldulensis and E. robusta.

Genogroup I noroviruses from five genetic clusters and genogroup II noroviruses from eight genetic clusters were detected in stool extracts using degenerate primers and single-tube, real-time reverse transcription-PCR (RT-PCR) with SYBR Green detection. Two degenerate primer sets, designated MON 431-433 and MON 432-434, were designed from consensus sequences from the major clusters of norovirus...