By identifying the sequence of retro- and lentiviral integration sites in peripheral blood leukocytes, the clonal composition and fate of genetically modified hematopoietic progenitor and stem cells could be mapped in vitro and in vivo. Previously available methods have been limited to the analysis of mono- or oligoclonal integration sites present in high copy numbers. Here, we perform characterization of multiple rare retroviral and lentiviral integration sites in highly complex DNA samples. The reliability of this method results from nontarget DNA removal via magnetic extension primer tag selection (EPTS) preceding solid-phase ligation-mediated PCR. EPTS/LM-PCR allowed the simultaneous direct genomic sequencing of multiple proviral LTR-flanking sequences of retro- and lentiviral vectors even if only 1 per 100 to 1000 cells contained the provirus. A primer walking "around" the integration locus demonstrated the adaptability of EPTS/LM-PCR to study unknown flanking DNA regions unrelated to proviruses. The technique is fast, inexpensive, and sensitive in minimal samples. It enables studies of retro- and lentiviral integration, viral vector tracking in gene therapy, insertional mutagenesis, transgene integration, and direct genomic sequencing that until now have been difficult or impossible to perform.

A comparison betweeen the electropherograms of the spelt and wheat cultivars showed specific differences in the gliadin band patterns which provided the possibility of a clear classification into spelt or wheat. A special nomenclature was developed to be able to improve the presentation of the gliadin band pattern of spelt, which is different from that of wheat. This nomenclature, however, has not yet been applied to other cereals. The gliadin band patterns were presented in a schematic form. As a parameter for comparison, idealized band patterns of both wheat and spelt were developed by comparing the proportions of the bands of all available types. When comparing the gliadin band patterns of the spelt cross-breeds with their corresponding parental generations, it was noted that the same parental bands were not always transmitted and that the cross-breeds showed differences in the intensity, mobility, occurrence, and the splitting of single bands. In general it can be said that the band pattern of the daughter generation - even in the examined F(5) and F(6) generations - is more similar to the band pattern of the mother than to that of the father, which proves a maternal effect.

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