TABLE 1

Summary of ASO markers

ASO markerCytological
position
PCR conditionaPCR primersASO probesbASO wash
conditionb,c
Pdp166A60°/55°CCTGTCGGAGAACAACATACCS: GTATAGCGGTTCTTG37°
AGGAGGCGAACGAAAATGM: CAATAGGTATTGGTA
trad73A56°GAACAAGCGAGAGGGATAGCS: GTGGATTATACCGAT50°
CTTTGGCGGTGGATTATACCM: ATCGGTACAATCCAC
Dbp73D73D55°TTAGTCGTCCTTCCTGTGGCAGAGS: GATCTTGTGACAATG47°
ATCAGCACCTTCTTCCGTTGCGTCM: GATCTTGAGACAATG
ash176B6-955°TTTCATCAGCAGTGGCGGTGS: TGAATTGTCTGACCA55°(S)/52°(M)
CGGCGTTTTGTAATGTCCCTGM: TGAATTGCCTGACCA
rdgCd77B63°CAAAGACATCGACTCAGCTACGS: CTCCACCACAGATAA50°
CGAACTCTCCACGATGCCM: TTATCTGCGGAGGAA
kni77E255°ACCAGACATGCAAAGTGTGCGGS: TGTATCTAAAACCTA45°
TTCGACATGCCCACGTTGTAGM: TAGGTTTCAGATACA
Pka77F1-555°AAAACCGATGTGAAGCTGTGGGGS: TGCTAAGACAAATAG40°
CGTGATGTTGCGTTTGAGTATGTCM: TGCAATGACAAATGG
Edg7878E53°CGAGCACTGTTGTTATTGTTGCCS: GGATGAGACCCTTGT57°(S)/52°(M)
TTCTGGAAGCTGCGGATCTGM: ACAAGGGACTCATCC
Rga83B55°GATGGTGAGTGCTGGAACACTTCS: CAACCTTCCTGTTAC45°(S)/37°(M)
GTTAATCAGATATTCGGGTGGCACM: CAACTTTCCTGGTAC
lbl93E55°CTTTCCGCTGCCGATATAGTTGS: CCTGTTATAAGGAGAA32°
ACGTCCTTCTTCAGCTCCTCCATGM: CCTGTTAAAGGAAAA
Hsr93D55°GAGGCAGTTATCCAGGATGTAAGGS: TTCATCGTGTTGCGT47°(S)/42°(M)
ATGATCTCAGTCACGGTCGCAGACM: TTCATCAATGTTGCGT
Fsh90C55°TGGCTGCGTATTTCCGTCTGS: AGAATAGATGAATCG50°(S)/47°(M)
CAACTTGATGCGATGGTTCAGGM: GCATTCAACTATTCT
Fas1d89E1-260°/55°GCGACTCTTTGAGTATCCGCS: TACCAATTTTCTATTG37°
GGCCAAGATGATGCAAATGM: CAATAGGTATTGGTA
Akt188B955°GTAATAGCAGCGGCGTTAAGAAAGS: CTTGTTAATAGTATT40°(M)/37°(S)
CGGGAATTTTACCTCCTCCACCM: CTTGTTACTAGTATT
Su(Hw)88B355°CAAGAACTCCCTCCAACAAGCAGS: TTTCCACACACGTGC50°(S)/45°(M)
CGCAAACGATACCCACTGTCATAGM: TTGCCACACACATGC
ems88A1-255°GCCCAGTTTATGCCCAATCCS: AAATTTTTGTCACTG42°(S)/37°(M)
CGATTCGCCAGGATAAGGAACM: AAATTTGAGTCTCAT
Mstd87F50°TCCTTTGCCTCTTCAGTCCGS: AGAGAAATGAAAATG37°
TCCACAGGCATAGCATGGTCM: CATTTCCATTTCTGT
Odhd86D55°CAGTGGTGTTGGGACATGAGS: GTGGGTTCGTGGAAC50°
TGGCCAGCTCAAACTTGTCM: GTTCCACAAACCCAC
Crc86BC55°AAAGTGGTGCGGGATCGTTCACS: TTACTATAGCTACTC37°
CATCCTGAGAGGTCTGAATGCCM: GAGTTGCTATCGTAA
tld96A55°GCCGAAGGAGCCAAAGGATGS: CAACTATTGAAGAGG45°
AAACCCGATTGTGTGACCCAGM: CCTCTTCCATAGTTG
  • In addition to 20 ASO markers listed here, 18 more had been developed before (Liu et al. 1996; Zeng et al. 2000). Their locations are 61A(Lsp1), 62A(ve), 64B-C(Acr64B), 65E(Dbi), 66D(h), 68E(CycA), 70D(fz), 71CD(Eip71CD), 82C(5-Ht2), 84B (Antp), 92B(ninaE), 85E(Tub85E), 85A3-B1(hb), 95D(Rox8), 97AB(Ald), 98B(Mlc1), 99D(jan), and 100E(Efld2).

  • a All PCR reactions followed this format: 94° for 3 min/1 cycle; (94° for 45 sec, annealing temperature for 30 sec, 72° for 15 sec)/40 cycles. If two annealing temperatures are given, the first was used for 10 cycles and the second for 30 cycles.

  • b S, D. simulans-specific; M, D. mauritiana-specific.

  • c 32P-labeled probes were washed at designated temperatures for 15 min.

  • d Also used in Liu et al. (1996) and Zeng et al. (2000), but ASO probes were newly developed.