and Implications The ELISA assay is a more rapid and sensitive method for PCR product detection than conventional gel electrophoresis. The PCR/ELISA allows for the processing of numerous samples, is relatively inexpensive and eliminates the need for electrophoretic and photographic equipment as well as the use of potential carcinogens such as ethidium bromide which is utilized in the gel electr...

One case of paediatric Ewing's sarcoma and two peripheral primitive neuroectodermal tumours/extra-osseous Ewing's sarcoma were studied for the characteristic t(11;22) translocation, using a recently described RNA-polymerase chain reaction method (RNA-PCR). PCR products of the expected sizes were obtained from RNA derived from the Ewing's sarcoma and the peripheral primitive neuroectodermal tumo...

In order to discern sake yeast, shochu yeast, and wine yeast, a specific polymerase chain reaction (PCR) was investigated. PCR products between shochu yeast strains and sake yeast weredistinguished by using a primer designed with an open reading frame of FLO5. Wine yeasts and most of other yeasts were distinguished by a primer designed with an open reading frame of YHR213W.However, PCR products...

Here we define a new approach for the detection and characterisation of Leishmania complexes by polymerase chain reaction (PCR) and specific hybridisation. The first step consists of PCR amplification of kDNA minicircles using general kinetoplastid primers, which generate a polymorphic multi-banding pattern for all Leishmania species and other Kinetoplastidae. The second step is the identificat...

Construction of a random ssDNA sublibrary is an important step of the aptamer screening process. The available construction methods include asymmetric PCR, biotin-streptavidin separation, and lambda exonuclease digestions, in which PCR amplification is a key step. The main drawback of PCR amplification is overamplification increasing nonspecific hybridization among different products and by-pro...

Quantitative PCR can often be improved by conducting the amplification with nested primers. First, fewer nonspecific amplification products, which could otherwise interfere with quantitation, are produced. Often, nonspecific products can be eliminated. In these cases, relatively simple nonspecific detection techniques are suitable for quantitation. In addition, nested primer PCR provides intrin...

Figure S1. Additional validations of microarray predicted ESRP regulated splicing events. Semiquantitative RT-PCR was used to assay a set of more complex candidate alternative splicing events. Identities of PCR products are labeled to the right of the gels. The percent inclusion of the relevant exon(s), shaded in red, is given below each lane. For the mutually exclusive splicing events in the F...

We present a method for performing highly parallel PCR reactions in a picowell array (PWA) simultaneously immobilizing generated PCR products in a covalent and spatially-resolved manner onto a microscope slide via solid-phase PCR (SP-PCR). This so called PWA-SP-PCR was performed in picowell arrays featuring 100,000 wells cm(-2) of 19 pL reaction volumes with a surface-to-volume ratio of 0.2 μm(...

BACKGROUND The applicability of microarray-based transcriptome massive analysis is often limited by the need for large amounts of high-quality RNA. RNA arbitrarily primed PCR (RAP-PCR) is an unbiased fingerprinting PCR technique that reduces both the amount of initial material needed and the complexity of the transcriptome. The aim of this study was to evaluate the feasibility of using hybridiz...

Immobilized Metal Affinity Chromatography (IMAC) has been used for decades to purify proteins on the basis of amino acid content, especially surface-exposed histidines and "histidine tags" genetically added to recombinant proteins. We and others have extended the use of IMAC to purification of nucleic acids via interactions with the nucleotide bases, especially purines, of single-stranded RNA a...