After establishing haplotypes for the family, multiplex PCR reactions were optimized on single
lymphocytes isolated from family members as well as from consenting donors. This was done with careful
manipulation of individual primer concentrations and optimization of the PCR annealing temperature,
until PCR efficiency and Allele Dropout (ADO) levels were at satisfactory levels (94% and 7% respectively).
The optimized protocol was used in the PGD case.