Table 1 PCR amplification results for 25 cytologically karyotyped males.

♂♂	Cytological Karyotype		PCR fragments¹									Molecular Karyotype
♂♂	Cytological Karyotype		AB	AH2	I C	AK	AG	KL	CD	AL	H1H2	Molecular Karyotype
1 M	E_st	E_st	+					+	+		+	E_st	E_st
2 M	E_st	E_st	+					+	+		+	E_st	E_st
3 M	E₁₊₂	E_1+2+9+12			+	+	+ +	+	+	+	+	E₁₊₂	E_1+2+9+12
4 M	E_st	E₁₊₂	+				+	+	+		+	E_st	E₁₊₂
5 M	E_st	E_1+2+9+12	+		+	+	+	+	+	+	+ +	E_st	E_1+2+9+12
6 M	E₁₊₂₊₉	E_1+2+9+12			+	+	+		+	+	+	E₁₊₂₊₉	E_1+2+9+12
7 M	E_st	E₁₊₂	+				+	+	+		+	E_st	E₁₊₂
8 M	E_st	E_st	+					+	+		+	E_st	E_st
9 M	E₁₊₂	E_1+2+9+3		+			+	+	+	+	+	E₁₊₂	E_1+2+9+3
10 M	E_st	E_st	+					+	+		+	E_st	E_st
11 M	E_st	E_st	+					+	+		+	E_st	E_st
12 M	E_1+2+9+12	E_1+2+9+12			+ +	+	+			+	+	E_1+2+9+12	E_1+2+9+12
13 M	E₁₊₂	E_1+2+9+12			+	+	+	+	+	+	+	E₁₊₂	E_1+2+9+12
14 M	E_st	E₁₊₂	+				+	+	+		+	E_st	E₁₊₂
15 M	E₁₊₂₊₉	E_1+2+9+12			+	+	+		+	+ +	+	E₁₊₂₊₉	E_1+2+9+12
16 M	E_st	E_1+2+9+12	+		+	+	+	+	+	+	+	E_st	E_1+2+9+12
17 M	E_st	E_1+2+9+3	+	+			+	+	+	+	+	E_st	E_1+2+9+3
18 M	E₁₊₂	E₁₊₂₊₉	+			+	+	+	+	+	+	E₁₊₂	E₁₊₂₊₉
19 M	E_st	E₁₊₂₊₉	+		+	+	+	+	+	+	+	E_st	E₁₊₂₊₉
20 M	E_st	E_1+2+9+12			+	+	+	+	+	+	+	E_st	E_1+2+9+12
21 M	E_1+2+9+12	??			+	+	+	+	+	+	+	E₁₊₂	E_1+2+9+12
22 M	E_1+2+9+12	E_1+2+9+12	+				+			++	+	E_1+2+9+12	E_1+2+9+12
23 M	E_st	E₁₊₂₊₉	+				+	+	+	+	+	E_st	E₁₊₂₊₉
24 M	E_st	E₁₊₂₊₉	+				+	+	+	+	+	E_st	E₁₊₂₊₉
25 M	E_st	E₁₊₂					+	+	+		+	E_st	E₁₊₂

The karyotype of 24 of the 25 males collected in November 2014 could be cytologically established whereas for male 21 M, only one of its E chromosome arrangements could be identified. Fragments AB, AH2 and IC are exclusive for arrangements E_st, E_1+2+9+3 and E_1+2+9+12, respectively. +, a single amplification product; ++, two differently sized amplification products.
¹Initally, the amplification of seven PCR fragments (AB, AH2, IC, AK, AG, KL and CD) was tested in the 25 males sample. The relatively high dropout frequency of fragment AK detected in the 2015 sample led to its replacement by fragment AL, which required the additional amplification of fragment H1H2 to discern between the low-frequency karyotypes E₁₊₂₊₉/E_1+2+9+3 and E_1+2+9+3/E_1+2+9+3 (see text).

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