To get a pure genomic background, the knock-in lines were crossed with wild-type C57BL6 mice for seven further generations.
Her father, uncle and grandmother were affected, and four further generations were reported to be affected.
The F2 heterozygous (tardbp fh301/+) animals were bred with AB fish for three further generations to obtain F5 mutant heterozygotes.
In order to expand the population, the F9 Tir1 homozygous lines were then intermated for three further generations to produce 600 and 100 of D17SS and D17RR F12 mice respectively.
These lines have undergone three to four further generations of inbreeding than the lines described by Michaelson-Yeates et al. [ 14], and consequently, we started out from an assumption that the residual heterozygosity reported earlier [ 4] would be diminished.
Thus, while three to four further generations of inbreeding are likely to have increased the extent of homozygosity, a residual heterozygosity level of <1% for J5 and SC is unlikely.
Sir David King, Tony Blair's chief scientist, recently argued that one further generation of nuclear power stations is needed (in Britain at least) to buy time, in order to keep down emissions of carbon dioxide, the chief greenhouse gas, while new carbon-free non-nuclear technologies are developed.
We subsequently performed Illumina sequencing on three further white clover samples, generating 14 million transcript reads from a mixed sample comprising 24 divergent white clover genotypes, and 50 million reads on two further eighth generation white clover inbred lines.
Two further next-generation CYP17A1 inhibitors, VT-464 (Toren et al, 2015) and galeterone TOK-0011) (Handratta et al, 2005) are currently undergoing phase I and II development, respectively (NCT02012920; NCTable9734) (Table 1).
Further to this, preliminary investigations involving backcrossing for two generations to two further strains (C3HHeH and 129Svev) suggest that the phenotype remains penetrant (albeit incompletely) on other genetic backgrounds (data not shown).
Two further improvements for better signal generation is the use of VIC (a proprietary dye by ABI) as reporter dye yielding higher fluorescence than TET and the use of a dark quencher emitting transferred energy as heat (instead as fluorescence like TAMRA) and thereby reducing cross fluorescence with reporter dyes.
