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Quantitative PCR: the determination of template copy numbers by temperature gradient gel electrophoresis (TGGE) Low copy numbers of DNA or RNA sequences can be quantified easily by combining PCR (Polymerase Chain Reaction) and TGGE (Temperature Gradient Gel Electrophoresis). This combination allows to overcome the problems of quantitive PCR caused by plateau effects, uneven priming or, variable...

Escherichia coli and other Proteobacteria are augmented and several other bacteria are diminished in Crohn’s (CD) disease patients’ intestine. This imbalance in bacterial species composition—termed dysbiosis—seems to be determinant of CD manifestation. Since a great part of intestinal bacteria are not cultivable, detection of CD dysbiosis is accomplished by molecular tools, involving sequences ...

Strains belonging to the Gram-positive coryneform soil bacteria were screened genotypically by temperature-gradient gel electrophoresis (TGGE). This method allows the sequence-specific separation of amplified fragments of 16S rRNA genes. A total of 115 reference strains representing the majority of the species of the genera Aeromicrobium, Agromyces, Arthrobacter, Aureobacterium, Cellulomonas, C...

Hepatitis C virus (HCV) forms complex quasispecies populations which consist of a large number of closely related genetic variants. This genetic heterogeneity may cause antigenic variation or drug resistance. We used heteroduplex analysis by temperature gradient gel electrophoresis (TGGE) to characterize genetic variants of HCV. The high resolution of TGGE was proven by comparison of DNA sequen...

conclusions our successful extraction of purified lps from e. coli isolates of uti patients’ urine samples can be an important step to understand the uti disease conditions. results the silver-stained gel demonstrated both smooth and rough type lps by showing trail-like band patterns with the presence and lacking o-antigen region, respectively. coomassie blue staining showed no band assuring th...

The main bacteria in peaty, acid grassland soils in the Netherlands were investigated by ribosome isolation, temperature gradient gel electrophoresis, hybridization, cloning, and sequencing. Instead of using only 16S rDNA to determine the sequences present, we focused on rRNA to classify and quantify the most active bacteria. After direct ribosome isolation from soil, a partial amplicon of bact...