Table 1: Test and reference conditions for the optimisation experiments.

ConditionPreparatory workMultiplex oligonucleotide ligationPCRHybridisation and analysis on MAGPIX
DNA isolationa   Probe concentration
(nM)
 DNA
 volume
(µL)
Taq DNA ligase (units)  Initial denaturation
(min)
Ligation product volume
(µL)
Taq DNA polymerase
(units)
   Cycles PCR product volume (µL) Microspheres per
reaction
SAPE concentration
(µg/mL)
Reporter dye

Experiment30010050IGM12524246510350.250.535402.55all3507502500 4 10SAPEAlexa 532

1Rnslslsxxxxxxxxx xx
2xgsRgsxxxxxxxx xx
3xxRgsRgsgsxxxxxx xx
4xxxxRlsxxxxx xx
5xxxxxRnsxxxx xx
6xxxxxxRgsxxx xx
7xxxxxxxRgsxx xx
8xxxxxxxxRlslsx xx
9xxxxxxxxxlslsRls Rx
10xxxxxxxxxx xRgs

R: reference condition; x: condition used in experiment; ns: test condition which is not significant at in two-sided hypothesis test; ls: test condition which is significant at in one-sided hypothesis test with alternative hypothesis true location shift is less than 0; gs: test condition which is significant at in one-sided hypothesis test with alternative hypothesis true location shift is greater than 0; bold type: optimal condition as concluded from the experiment.
a300: single colony in 300 µL sterile deionised water; 100: single colony in 100 µL sterile deionised water; 50: single colony in 50 µL sterile deionised water; IGM: InstaGene Matrix.