There were three cases in the category reported to have normal karyotype, normal locus specific FISH test at the DGS/VCFS critical region and normal telomeric FISH test.
The aim of the strategy was to insert a mutant tub2 gene copy (either SA or SE) into its normal locus (see Materials and Methods, and Figure 1A for partial amino-acid sequence of mutants).
These data could justify that tumors with AIs, and thus carrying a likely deletion at this locus, may have a worse prognosis than tumors with a normal locus.
We found that Exo1-4S∷E was less functional than Exo1 when expressed from the normal locus because cdc13-1 exo1-4S∷E mutants formed larger colonies than cdc13-1 EXO1 stratns at 26 and 26.5°C.
Although HR has the important advantage that elements are assessed in the normal context, HR carries the disadvantage that the normal locus sometimes contains redundant or counteracting elements that obfuscate analysis.
A modified version of the method by Schuelke (2000) was used, employing three primers in the PCR reaction: the fluorescent-labeled universal primer, a locus-specific forward primer with a universal tail attached at its 5′ end, and a normal locus-specific reverse primer.
Thus, PCR reactions were performed with three primers: one primer of the microsatellite primer pair was designed with a universal M13 tail attached at its 5' end, the second primer was a normal locus-specific reverse primer, and the third primer was a fluorescently labeled M13 primer.
Normal ETV1 loci are visualised in interphase nuclei as immediately adjacent green and red signals.
This estimation is only possible when the BAFs of the tumour sample at normal heterozygous loci are available, DOC ratios alone would not allow correcting for this.
