Table 4.

Details of the 12 species tested and a summary of utility of the Conserved Avian Microsatellite (CAM) markers*

Species	Status	Sample type and storage	Gen dist to ZF (ΔT m H)	Gen dist to CH (ΔT m H)	Order & Family ([[25]] / NCBI Taxonomy Database)	PCR profile	Pop	Loci amp. (%)	Loci poly. (%)	Geno-typer	Samples taken and DNA extracted by	Sample supplier(s)
NEOGNATHAE
Passerines
Zebra finch	Captive	T/E &	0	28	Passeriformes	56	1	100	92	ADB	Jayne Pellatt,	Tim Birkhead
Taeniopygia guttata		B/E			Passeridae/Estrildidae						Jon Chittock
Berthelot’s pipit	Wild	B/E	8.3	28	Passeriformes	56	4	96	70	LGS	LGS	David Richardson,
Anthus berthelotii					Passeridae							Juan Carlos Illera
House sparrow	Wild	B/E	8.5	28	Passeriformes	56	1	96	78	ADB	Nancy Ockendon	TB
Passer domesticus					Passeridae
Chaffinch	Wild	B/E	10.0	28	Passeriformes	TD1	1	96	83	JP	Ben Sheldon	Ben Sheldon
Fringilla coelebs					Fringillidae
Eurasian bullfinch	Wild	B/E	10.0	28	Passeriformes	TD1	1	96	65	JP	Kate Durrant,	Tim Birkhead
Pyrrhula pyrrhula					Fringillidae						Stuart Sharp, Simone Immler
Great tit	Wild	B/E	11.1	28	Passeriformes	TD1	1	96	56	JP	Louise Gentle,	TB
Parus major					Paridae						Harrie Bickle
European blackbird	Wild	B/E	11.7	28	Passeriformes	TD1	1	83	60	JP	Michelle Simeoni	Ben Hatchwell
Turdus merula					Muscicapidae/Turdidae
Rifleman	Wild	B/E	19.7	28	Passeriformes	56	1	96	61	SAJP	SAJP	Ben Hatchwell
Acanthisitta chloris					Acanthisittidae
Non-passerines
Leach’s storm-petrel	Wild	B/E	21.6	28	Ciconiiformes	56	4	96	56	AWJB	AWJB	AWJB
Oceanodroma leucorhoa					Procellariidae
Barn owl	Wild	B/E	22.5	28	Strigiformes	TD1	1	92	32	JP	Akos Klein	Akos Klein
Tyto alba					Tytonidae
European roller	Wild	B/E	25.0	28	Coraciiformes	B,	1	96	39	DM-G,	DM-G	Deseada Parejo,
Coracias garrulus					Coraciidae	TD2				MM-M		Jesus Avilés
PALAEOGNATHAE
Chicken (domestic)	Captive	B/E	28.0	0	Galliformes	TD1	1	100	38	JP	Hans Cheng	Hans Cheng
Gallus gallus domesticus					Phasianidae

*Four individuals were tested per species with 24 Conserved Avian Microsatellite (CAM) primer sets. All PCR failures were rechecked for amplification by a different researcher (GJH) using the touchdown PCR program (TD1);

PCR profiles:

A: QIAGEN Multiplex PCR Master Mix; 95°C for 15 minutes, followed by 35 cycles of 94°C for 30 seconds, 56°C for 90 seconds, 72°C for 1 minute, and finally 60°C for 30 minutes.

B: (used only for the unrelated rollers), Bioline DNA Taq polymerase, 94°C 3 min, then 35 cycles of 94°C for 30 s, 56°C for 30 s, 72°C for 30 s, and finally 72°C for 10 min.

TD1: QIAGEN Multiplex PCR Master Mix; touchdown PCR program, 95°C for 15 min followed by 16 cycles of 94°C for 30 s, 65°C for 90 s decreasing by 1°C per cycle, 72°C for 60 s for 10 cycles, followed by 94°C for 30 s, 55°C for 90 s, 72°C for 60 for 25 cycles, with a final step of 72°C for 10 min.

TD2: (used only for the related European rollers) Bioline DNA Taq polymerase, touchdown PCR profile, 94°C for 3 min, then 10 cycles of 94°C for 30 s, 65°C for 30 s (and decreasing by 1°C for 15 cycles), 72°C for 1 min, followed by 28 cycles of 94°C for 30 s, 50°C for 30 s and 72°C for 30 s, followed by one cycle of 5 min at 72°C.

T, tissue; B, blood; E, ethanol; Pop., number of populations represented in the four individuals tested; amp., amplifying; poly., polymorphic; Loci poly. (%) indicates the proportion of loci polymorphic of those amplifying.

Genetic distance to ZF, genetic distance from species tested to zebra finch based on [33] and the classification of [25]; Genetic distance to CH, genetic distance from species tested to chicken [33].