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Biomedical subjects

M P Starkey

Publications and source records attributed to M P Starkey.

7 recordsLinked to original sources
PubMed1998

A simple and efficient method for the isolation of differentially expressed genes.

A simple and reproducible general approach for the isolation of differentially expressed genes is described. Digestion of cDNAs with a class IIs restriction endonuclease produces fragments with every combination of possible bases in the cohesive ends. Under stringent conditions, the specific ligation of adaptors with perfectly complementary overhangs partitions the cDNA fragments into non-overlapping subpopulations. Internal cDNA restriction fragments are exponentially amplified by adaptor primer PCR and visualised by non-denaturing polyacrylamide gel electrophoresis. The power of the technology was demonstrated using a rat model of pressure-induced left-ventricular hypertrophy (LVH). A set of 29 fragments, derived from a sample (6 %) of the possible adaptor pool combinations, displayed apparent differential expression. The differential expression of 19 (66 %) were confirmed by Northern blot analysis. Sequence analysis identified both genes known to be upregulated in LVH, and novel genes. The fidelity of adaptor ligation was demonstrated by the isolation of known gene fragments by appropriate adaptor combinations. The spiking of mRNA populations with known amounts of a synthetic mRNA demonstrated a current sensitivity equivalent to the detection of transcripts expressed at the level of as little as 1 in 10,000 molecules.

Animals↗
PubMed1998

Reference cDNA library facilities available from European sources.

cDNA libraries are the cornerstone of efforts to identify the relatively small regions of genomes that are responsible for biological effects. Gene hunter seeking candidate genes, via a variety of approaches, ultimately focus on the cloning, sequencing, and expression of cDNAs. Assistance is now available to researchers in the form of genome programs, whose initial goals include assembly of a complete collection of expressed sequences derived from the genome of interest. The concept of reference sets of cDNA libraries is that the aims of genome programs are served most effectively by different laboratories working on a common set of high-quality arrayed cDNA libraries, using different experimental approaches, thereby reducing unnecessary duplication of effort, and maximizing the amount of information that one set of resources can provide.

DNA, Complementary↗
PubMed1992

The specialisation of oral surgery in general practice.

Much has been written recently on the subject of introducing a specialist title in oral surgery and the establishment of specialist oral surgery practices. The aim of my elective was to conduct a literature review, interview general practitioners involved in oral surgery, and examine the feasibility of undertaking oral surgery in a specialist practice.

Anesthesia, Dental↗
PubMed1991

Design and use of synthetic oligonucleotide probes in the cloning of delta-endotoxin genes from Bacillus thuringiensis.

A detailed protocol is described for the design and use of synthetic oligonucleotide probes for screening DNA libraries from Bacillus thuringiensis var. kurstaki (strain HD191) for copies of the gene (tox) encoding the insecticidal delta-endotoxin. Two homologous tox genes were identified in this organism; one of these was located on a 75-kb plasmid and the other on a second large plasmid or the bacterial chromosome. A tox gene was isolated as a 6.5-kb HindIII fragment of B. thuringiensis plasmid DNA.

Bacillus thuringiensis↗