A PCR-based method for isolation of genomic DNA flanking a known DNA sequence

We describe a simple PCR-based method for the isolation of genomic DNA that lies adjacent to a known DNA sequence. The method is based on the directional cloning of digested genomic DNA into the multiple cloning site of a pUC-based plasmid to generate a limited genomic library. The library is plated onto a number of selective LA plates which are incubated overnight, and recombinant plasmid DNA is then isolated from resistant colonies pooled from each plate. PCR amplification is performed on the pooled recombinant plasmid DNAs using primers specific for the pUC vector and the known genomic sequence. The combination of efficient directional cloning and bacterial transformation gives relative enrichment for the genomic sequence of interest and generates a simple DNA template, enabling easy amplification by PCR

Rapid localization of proteins in chromatographic eluates

Rapid localization of proteins in chromatographic eluate

The separation of malic dehydrogenase isoenzymes of Neurospora crassa by polyacrylamide gel electrophoresis

The separation of malic dehydrogenase isoenzymes of Neurospora crassa by polyacrylamide gel electrophoresi

Avoid adding insult to injury - correct management of sick female endurance athletes

Objectives. To evaluate the efficacy of Ringer’s lactate, isotonic saline and hypertonic saline on the clinical and biochemical recovery of athletes with exercise-associated hyponatraemic encephalopathy caused by fluid overload. Methods. We retrospectively reviewed serial blood sodium concentrations (Na+) and qualitative signs of recovery and time to recovery in two healthy menstruant females hospitalised with dilutional exercise-associated hyponatraemic encephalopathy after withdrawal from the 2011 Comrades Marathon (89 km) and Argus Cycle Tour (109 km). Results. Improvements in blood Na+ did not occur with intravenous administration of Ringer’s lactate solution, but did occur with administration of isotonic and hypertonic saline. Qualitative improvements in mental status were not quantitatively related to the biochemical value of blood Na+ or subsequent return to normonatraemia. Conclusions. Hyponatraemia should be suspected in all female athletes presenting to the medical area of endurance races with vomiting, altered mental status and a history of high fluid intake. If a diagnosis of exercise-associated hyponatraemia with cerebral encephalopathy is confirmed, the treatment of choice is administration of an intravenous bolus of hypertonic saline. Administration of Ringer’s lactate should be discouraged, as this does not correct Na+ and appears to delay recovery