Next-generation sequencing platforms are powerful technologies, providing gigabases of genetic information in a single run. An important prerequisite for high-throughput DNA sequencing is the development of robust and cost-effective preprocessing protocols for DNA sample library construction. Here we report the development of a semi-automated sample preparation protocol to produce adaptor-ligat...

Differential display reverse transcription PCR (DDRT-PCR) is a procedure used to identify the induction or repression of gene expression. In most DDRT-PCR protocols, radioisotopes are incorporated during PCR and the cDNA products are detected by autoradiography. This report describes the fluorescent labeling of cDNAs and their detection on automated sequencers from PE Applied Biosystems. A fluo...

To facilitate evolutionary and forensic studies of DNA polymorphisms on the Y chromosome, we devised a multiplex amplification procedure for short-tandem-repeat (STR) loci. Four tetranucleotide STR loci (DYS19, DYS390, DYS391, and DYS393) were simultaneously amplified with FAM-labeled primers and genotypes were determined with an automated DNA sequencer. We typed 162 males from three U.S. popul...

A variant of the cDNA-AFLP method coupled to an automated sequencer was used to quantify transcripts differentially expressed between sexes of the marine snail Littorina saxatilis. First, we conducted a validation study of the technique using known concentrations of a commercial marker. Second, we analysed six replicates of males and females from a population showing no apparent sexual dimorphi...

A major challenge of the Human Genome Initiative is the development of a rapid, accurate, and efficient DNA sequencing technology. A major limitation of current technology is the relatively long time required to perform the gel electrophoretic separations of DNA fragments produced in the sequencing reactions. We demonstrate here that it is possible to increase the speed of sequence analysis by ...

Terminal deoxynucleotidyltransferase (terminal transferase, E.C. 2.7.7.31) has been used to add a single fluorescent ribonucleotide to the 3' terminus of DNA sequencing primers, thereby creating primers suitable for automated DNA sequence analysis. The previously introduced procedure using fluorescein-UTP for the postsynthetic labeling of primers can, under appropriate reaction conditions, now ...

The massive scale of DNA sequencing for the Human Genome Initiative compels efforts to reduce the cost and increase the throughput of DNA sequencing technology. Contemporary automated DNA sequencing systems do not yet meet estimated performance requirements for cost-effective and timely completion of this project. Greater accuracy of basecalling software would minimize manual review and editing...

background: breast cancer is the most common malignancy in iranian women. mutations in brca1 gene is one of the important genetic predisposing factors in breast cancer. this gene is a tumor suppressor that plays an important role in regulating the functions of rad51 protein for strand invasion in homologous recombination repair. methods: the brca1 gene has amplified in the dna isolated from bre...

This paper describes an interactive multi-track MIDI sequencer for generating polyrhythmic patterns. Patterns are determined by the state of a set of autonomous Boolean networks, each node of which corresponds to a single musical event. This event will be triggered if the Boolean node is active and the sequencer step corresponds to its location in the pattern. Users may alter the networks which...