Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society 

About: Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society is an academic journal. The journal publishes majorly in the area(s): Gel electrophoresis & Agarose. It has an ISSN identifier of 0954-6642. Over the lifetime, 145 publications have been published receiving 1366 citations.

Modifications to improve the effectiveness of restriction fragment length polymorphism typing.

Abstract: A streamlined and effective method for RFLP analysis of DNA has been developed. Southern transfers are accomplished by alkali blotting DNA onto positively charged nylon membranes. The prehybridization step has been eliminated. The hybridization solution is composed of three cost-effective reagents: 7% SDS, 10% PEG, and phosphate buffer. By using probes that hybridize to variable number of tandem repeat loci, and RFLP analysis of one to two micrograms of genomic DNA can be achieved within five working days under normal working hours. With longer autoradiographic exposures, as little as 20-100 ng of human genomic DNA is sufficient for analysis.

Cerebrospinal fluid protein variations in common to Alzheimer's disease and schizophrenia.

Abstract: Analysis of silver stained two-dimensional (2D) gels of cerebrospinal fluid (CSF) from 27 patients with schizophrenia (SCZ) and 10 patients with Alzheimer's disease (AD) revealed an increase in the relative amount of a polypeptide of 18,000M(r) and isoelectric point of 6.5 when compared to the appropriate controls. This protein was identified by its electrophoretic characteristics and by immune analysis of Western blots as an isoform of alpha-2 haptoglobin, provisionally identified as alpha-2FS haptoglobin. Alzheimer's disease versus control CSF samples showed a 6.8-fold increase in the percent mean density value of this haptoglobin isoform (n = 10 AD vs 11 control; P > 0.025) while a 4.4-fold increase was observed in the schizophrenic patients (n = 17 SCZ vs 10 control; P > 0.001). Two additional polypeptides (proteins '127' and '128') of 40,000 M(r) and isoelectric points 5.7 and 5.9, respectively, described previously by this laboratory, were found in the CSF of 27% of schizophrenics, 23% of the Alzheimer's disease patients, and 4% of the controls in the current study. The presence of proteins 127 and 128, as well as the increased concentrations of alpha-2 haptoglobin in the CSF of Alzheimer's disease and schizophrenic patients, may be useful as diagnostic biological markers. They may also indicate a common pathophysiology between these diseases.

'Catalysts' for polyacrylamide gel polymerization and detection of proteins by silver staining.

Abstract: The crosslinker diacrylyl-piperazine produces polyacrylamide gels which display improved electrophoretic separation of proteins and better physical strength. It also produces gels with improved detection of proteins by ammoniacal silver staining by reducing the background. This reduced background provided us with an opportunity to investigate residual background staining caused by the catalytic reagents utilized in the polymerization of acrylamide gels. The commonly used catalyst system, tetramethyl-ethylenediamine and ammonium persulfate was shown to be responsible for the yellow staining background found after a prolonged development time with silver staining. An alternate catalyst system has been designed to decrease further the formation of this background staining. Dimethyl-piperazine or tetramethylethylenediamine, potassium or ammonium persulfate, and sodium thiosulfate are shown to provide for gels which have excellent mechanical and staining characteristics. These catalytic systems produce little background staining despite prolonged development time with the ammoniacal silver stain, and they reduce background staining with the dichromate silver stain.

Use of image analysis to quantitate changes in form of mitochondrial DNA after x-irradiation.

Abstract: In order to develop a sensitive method that can reliably assess the amount of previous genotoxic exposure, we examined whether relative loss of the supercoiled form of mitochondrial DNA in tissue culture cells could retrospectively reflect exposure to x-irradiation of 375,750, and 1500 roentgens. Five routines of a graphic analysis program named IMAGE developed for the Apple Macintosh II computer were used to densitometrically quantitate the relative amounts of supercoiled and nicked-circular forms of mitochondrial DNA blotted onto nitrocellulose. After x-irradiation, there were dose-dependent losses of supercoiled relative to nicked-circular DNA forms. Further developments in the methodology to evaluate changes in form of mitochondrial DNA should increase the sensitivity and reliability of the dosimetry. The IMAGE program is useful for a wide variety of studies in molecular biology, and is available at no charge.

The use of fluorescence detection and internal lane standards to size PCR products automatically.

Abstract: We have developed chemical procedures, optical and electrophoretic instrumentation and computer software automate the analysis of polymerase chain reaction (PCR) products. DNA molecules labeled with up to four different fluorescent dyes are analyzed within a single electrophoresis gel lane. A size calibration curve is created for each electrophoresis lane from the electrophoretogram of uniquely labeled DNA fragments belonging to an internal lane standard that co-electrophoreses with the PCR products. The unknown molecular lengths of PCR products are automatically calculated from the calibration curve. Data from control experiments with DNA segments of known molecular length demonstrate the accuracy and precision of such sizing. This system has been applied to the analysis of PCR products for research in the areas of human identification, genetic mapping and genetic disease.