The notion of a limitation is broad and elastic, encompassing restrictions on such "basic" actions as lifting one's arm (Nordenfelt, 1997; compare Amundson, 1992: actions "at the level of the person"); on more complex physical activities such as dressing and toileting; and on social activities like working, learning or voting (see Wasserman, 2001).
Explanatory variables encompass restrictions to specified centres or specialists, previous approval by a medical advisor of the health insurer or the requirement for a registry.
The sequence encompassing different restriction sites on pGl3-Basic vector are given in Additional file 4: Figure S7.
Optimizing the TJ tightness of hCMEC/D3 cell monolayers still remains a major challenge in order to provide an in vitro model that might recapitulate all the characteristics of human BBB, encompassing permeability restriction with appropriate molecular exclusion and functional efflux and influx transport systems.
A randomly 32P-labeled (Rediprime II Random Prime Labelling System, Amersham Biosciences) restriction fragment encompassing all of exons 1 and 2, all of the ORF and a part of exon 3 (XbaI-fragment) was used as a PrP probe.
Upon initialization and depending on the depth of the HiC dataset, the software splits the contigs into smaller pieces/bins encompassing at least 2 restriction fragments.
To genotype the G894T polymorphism in the exon 7, we performed a polymerase chain reaction (PCR) amplification of 206 bp fragment encompassing the variant, followed by restriction fragment length polymorphism (RFLP) using the restriction endonuclease Mbo1 (Promega).
To search for additional parvovirus variants, we used the new NS1/7.5EC PCR assay whose primers were designed from a conserved region of the B19/V9 sequence and encompasses an MfeI restriction enzyme site that would allow differentiation between B19- and V9-like sequences.
The homozygous 17 nt deletion that we identified in SMIM1 from Vel− subjects encompasses a StyI restriction site (Fig 4B), which enabled us to develop a PCR-RFLP analysis to screen for this deletion (Fig 4C).
For the restriction analysis, cDNA encompassing the respective mutation site was amplified using mutation-specific primers (Online Resource).
