Table 1.

Characteristics and results of alpha satellite constructs tested for human artificial chromosome formation^c

Chromosome	Starting BACa	Assay construct							Human artificial chromosome assay
		Name	Higher-order repeat α-satellite			Monomeric α-satellite			No. of colonies tested	No. (%) of colonies with HACs	CENP-A IF-FISH	Mitotic stability
			Size (kb)	No. of CENP-B boxes	Presence of CENP-A enrichment	Size(s) (kb)	No. of CENP-B boxes	Presence of CENP-A enrichment
Xp	RP13-971O21	DXZ1-9HOR	19	26	+	59, 23b	1	−	19	4 (21)	2/2	2/2
Xp	RP11-65I6	X-monomeric	0	NA	NA	129	14	−	30	0	NA	NA
3p	RP11-557B13	3-Monomeric	0	NA	NA	203	22	−	24	0	NA	NA
4q	RP11-365H22	D4Z2bn-9HOR	23	37	−	9	0	−	15	4 (27)	2/2	1/1
7q	RP11-435D24	D7Z2-8HOR	22	53	−	27	2	−	18	5 (28)	4/4	2/2
11p	RP11-100E23	D11Z1-7HOR	14	9	−	27	8	−	17	4 (24)	4/4	2/2
17p	RP11-458D13	D17Z1B-7HOR	17	27	−	132	0	−	20	4 (20)	2/2	2/2
19p	RP11-21H14	D19Z1-10HOR	29	6	−	19	0	−	10	0	NA	NA

^aThe genomic location of each BAC is indicated in Fig. S3a to h in the supplemental material.

^bTwo versions of DXZ1-9HOR containing different amounts of monomeric α-satellite were tested. Results were indistinguishable and were pooled.

^cAlpha satellite arrays from different chromosomes and representing different types of arrays were chosen for their capacity to form de novo centromeres in human artificial chromosome assays. A 19-kb stretch of DXZ1, a CENP-A-associated array within the context of the HuRef genome, was chosen as a control for use of a relatively small amount of higher-order repeat (HOR) array DNA in this assay. Two monomeric arrays, from chromosomes Xp and 3p, were chosen due to their inclusion of CENP-B boxes and the absence of higher-order repeat array DNA. Five additional arrays were included to test the competency of nonfunctioning sequences in the HuRef genome. NA, not applicable.