Table 1.

D17S518 Results for Patient H3 to Demonstrate How Data Were Analyzed

A. D17S518 alleles (mutants in bold italics) present in small pools using normal control N380 PBLs, and PBLs and positive control tumor DNA from MSI-High HNPCC patient H3.
		No. of pools with the following alleles (bp):a
Identity	Genotype	<80	81	83	85	87	89	93	95	97	99	101	103
N380 PBL	89/93	0	0	0	0	0	116	116	0	0	0	0	0
H3 PBL	89/95	0	0	0	0	0	35	18	21	0	0	0	0
H3 TU	89/95	2	0	9	11	22	62	4	58	8	12	7	4

B. Calculation of patient H3 and normal control N380 mutant fragment frequencies (MF) in D17S518 small pools
Identity	Genotype	Exp.ge1	Est.ge2	#pools	Expected alleles3	Estimated alleles3	a14	a24	Vars5	MF6
N380 PBL	89/93	0.75	2.37	128	192	606	116	116	0	0.000
H3 PBL	89/95	0.75	0.34	128	192	87	35	21	18	0.302
H3 TU	89/95	0.75	1.19	128	192	306	62	58	79	0.711

Chromatograms for an experiment as in Figure 1 were analyzed and compiled into locus specific tables, as in A, showing the observed number of individual variants, bold , and progenitors and their sizes (GeneScore; www.hkasoftware.com/genescore.shtml). The data from all experiments at each locus were compiled into a table, B, which includes the g.e. that was expected and the total number of pools run.). The observed, estimated g.e. for all the data and the estimated total number of alleles seen were calculated. A mutant frequency (MF) and the significance of the MFs between sample and control were calculated from the MF and the bootstrap standard error calculated (SPPCRv.2; www.hkasoftware.com/sppcr.shtml).

^aNo alleles were observed at 91 bp.

¹Expected genome equivalents (exp. g.e.),

²estimated genome equivalents per pool (est. g.e.),

³expected and estimated total number of alleles analyzed at that locus (exp. alleles and est. alleles),

⁴numbers of progenitor allele 1 (a1) and allele 2 (a2),

⁵number of variants observed (Vars)

⁶estimated Mutant Frequency (MF). PBL, peripheral blood leukocyte.