Table S16. Primers and cDNA clones used for RNAi and in situ
hybridization. |
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sort |
aug3 gene ID |
NCBI gene ID |
plasmid vector |
cloning site |
method/resource (Accession
number) |
forward primer for cloning |
reverse primer for cloning |
primers for dsRNA synthesis* |
Use |
|
1 |
aug3.g4238 |
LOC107451717 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCGCCCAACTCTATACGACAGTCC |
GGAATTCCTCGGACTGAGTCTTCGATCTT |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
2 |
aug3.g4238 |
LOC107451717 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCGCCCAACTCTATACGACAGTCC |
GGAATTCGCGTATAGTATATCTCCACCAT |
T7KS - T7SK |
pRNAi
(specificity validation [#F1R3]) |
|
3 |
aug3.g4238 |
LOC107451717 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCACATGCAGACGTGTGCTCC |
GGAATTCTTTCAGCCGCTGAAGGCAC |
T7KS - T7SK |
pRNAi
(specificity validation [#F2R2]) |
|
4 |
aug3.g14287 |
LOC107440133 |
pBluescript II
SK(+) |
HindIII |
Pt-twist/pBS-km-twi#6-1HIII#2
(AB167807) [ref. 1] |
n.a. |
n.a. |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
5 |
aug3.g17735 |
LOC107444270 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCCATGGAACTCCTATTATCTGTC |
GGAATTCTTCATCTTTACCCTCTCCATCA |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
6 |
aug3.g17735 |
LOC107444270 |
pBluescript II
SK(+) |
BamHI/EcoRI |
PCR |
CGGGATCCCATGGAACTCCTATTATCTGTC |
GGAATTCTTCATCTTTACCCTCTCCATCA |
n.a. |
in situ
hybridization |
|
7 |
aug3.g16467 |
LOC107442263 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCCCCTAGCCTTTTCTCTTCTAGG |
GGAATTCAAGTTGCTGTAGTTGGCAATGT |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
8 |
aug3.g15167 |
LOC107441267 |
pTriplEx2 |
SfiI |
EST clone
(FY379644/eS7_SB_025_B01) [ref. 2] |
n.a. |
n.a. |
T7new -
T7_5TrpExLD2 |
pRNAi (pilot
screen) |
|
9 |
aug3.g132 |
LOC107445924 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCAAGGAAGTTCAGAAGCAGCAAG |
GGAATTCCCCACTCCTCATTCCTGAAATA |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
10 |
aug3.g5118 |
LOC110282806 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCCGTCCTTTAATTACAGCCAAGG |
GGAATTCAGCTCTGGTCAGTCTCGAAATC |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
11 |
aug3.g5118 |
LOC110282806 |
pBluescript II
SK(+) |
BamHI/EcoRI |
PCR |
CGGGATCCCGTCCTTTAATTACAGCCAAGG |
GGAATTCAGCTCTGGTCAGTCTCGAAATC |
n.a. |
in situ
hybridization |
|
12 |
aug3.g16893 |
LOC107442891 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCCGAGAAGAAAGCAGTCAAATCC |
GGAATTCTGCTTCTCTTGATGCATTTCCA |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
13 |
aug3.g3963 |
LOC107451332 |
pTriplEx2 |
SfiI |
EST clone
(FY375900/eS6_d1_51_E01) [ref. 2] |
n.a. |
n.a. |
T7new -
T7_5TrpExLD2 |
pRNAi (pilot
screen) |
|
14 |
aug3.g20850 |
LOC107449642 |
pTriplEx2 |
SfiI |
EST clone
(FY378818/eS7_SB_014_F12) [ref. 2] |
n.a. |
n.a. |
T7new -
T7_5TrpExLD2 |
pRNAi (pilot
screen) |
|
15 |
aug3.g7720 |
LOC107455388 |
pTriplEx2 |
SfiI |
EST clone
(FY374484/eS6_d1_33_H05) [ref. 2] |
n.a. |
n.a. |
T7new -
T7_5TrpExLD2 |
pRNAi (pilot
screen) |
|
16 |
aug3.g7720 |
LOC107455388 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCGTACATAAAACTGCCACACGTC |
GGAATTCTACTTGTACCCCTGGAGTTCTG |
T7KS - T7SK |
pRNAi
(specificity validation [#F1R1]) |
|
17 |
aug3.g7720 |
LOC107455388 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCGATTTTGTTCCTGTCAAAGAGC |
GGAATTCCGACAATCAGGATGTCTCATAT |
T7KS - T7SK |
pRNAi
(specificity validation [#F2R2]) |
|
18 |
aug3.g7720 |
LOC107455388 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCGAAAGTGCCCAACGTGGTATTG |
GGAATTCTCATAGGATGGTACTTCGATCA |
T7KS - T7SK |
pRNAi
(specificity validation [#F3R3]) |
|
19 |
aug3.g7876 |
LOC107455577 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCGTTCGAATACGTTGATGGCATA |
GGAATTCGCTGGGAAAGACAGCAAAATAC |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
20 |
aug3.g1424 |
LOC107446703 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCACTGATGGTAACTGCGACAATG |
GGAATTCGATGGTTCCGTCAATAATGGAT |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
21 |
aug3.g22959 |
LOC107436727 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCAATGACTCCAAGGGAGTTTGAA |
GGAATTCGCTTTGCTTCGTTATATTTGCC |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
22 |
aug3.g4633 |
LOC107452245 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCACGGTTGATGGGTGATTTTAAG |
GGAATTCTTGCAACACTTTCCATCCTAGA |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
23 |
aug3.g26874 |
LOC107439682 |
pTriplEx2 |
SfiI |
fuchi / EST clone
(LC671429/S7_d1_24_G06) [ref. 2] |
n.a. |
n.a. |
T7new -
T7_5TrpExLD2 |
pRNAi (pilot
screen) |
|
24 |
aug3.g26874 |
LOC107439682 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCGACATGTTCTCACCTATTTCCG |
GGAATTCTGTAATTGCCTTAAGTCCGATG |
T7KS - T7SK |
pRNAi
(specificity validation [#F1R1]; phenotype analyses; RNA-seq; ATAC-seq) |
|
25 |
aug3.g26874 |
LOC107439682 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCGGATTCGTCTAATGAAAGTGG |
GGAATTCCTTGATTTCGAGCATTGGCGAT |
T7KS - T7SK |
pRNAi
(specificity validation [#F2R4]) |
|
26 |
aug3.g26874 |
LOC107439682 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCGTTACACAAGAGGAAGCGACCA |
GGAATTCCATCCTGTAAAATGACTGAACG |
T7KS - T7SK |
pRNAi
(specificity validation [#F3R3]) |
|
27 |
aug3.g18041 |
none |
pTriplEx2 |
SfiI |
EST clone
(FY370001/S7_d1_20_F12) [ref.2] |
n.a. |
n.a. |
T7new -
T7_5TrpExLD2 |
pRNAi (pilot
screen) |
|
28 |
aug3.g1904 |
LOC107447737 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCAATGCATTAGTGTCCATGTTGG |
GGAATTCCTTCATCGCCTTCCATAGAATC |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
29 |
aug3.g4752 |
LOC107452419 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCAACCATTCCAATGTGAAGTGTG |
GGAATTCTTGTTGTCTGCTTTTATGTCCG |
T7KS - T7SK |
pRNAi (pilot
screen) |
|
30 |
aug3.g1125 |
LOC107445669 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCATGGCTGCTGTTTCAGAGGATA |
GGAATTCAGGTCTTTGATTATGTTGCTGC |
n.a. |
in situ
hybridization |
|
31 |
aug3.g2114 |
LOC107448182 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCAGTGACACTCAGGATCTGTGTC |
GGAATTCCGTCTTGCACGTCTTCATCGTC |
n.a. |
in situ
hybridization |
|
32 |
aug3.g20850 |
LOC107449642 |
pTriplEx2 |
SfiI |
EST clone
(FY378818/eS7_SB_014_F12) [ref. 2] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
33 |
aug3.g17950 |
LOC107444566 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCAGAGAGGGTCTCAGAAGTTAAG |
GGAATTCTAGTTCGAATTGAACACAGCTC |
n.a. |
in situ
hybridization |
|
34 |
aug3.g12453 |
LOC107437591 |
pZL1 |
SalI/NotI |
EST clone
(FY223452/At_eW_022_H09) [ref. 3] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
35 |
aug3.g27539 |
none |
pTriplEx2 |
SfiI |
EST clone
(FY380287/eS7_SB_032_D11) [ref. 2] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
36 |
aug3.g27086 |
LOC110283546 |
pBluescript II
SK(+) |
EcoRI |
PCR |
CGGAATTCAGAAGAATGGAGAAGGTC |
GGAATTCTCAACACCACAGGGTAGTTGGA |
n.a. |
in situ
hybridization |
|
37 |
aug3.g486 |
LOC107443634 |
pZL1 |
SalI/NotI |
EST clone
(FY217041/At_eW_002_A15) [ref. 3] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
38 |
aug3.g26914 |
LOC107439896 |
pTriplEx2 |
SfiI |
EST clone
(FY373480/eS6_d1_21_D01) [ref. 2] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
39 |
aug3.g132 |
LOC107445924 |
pBluescript II
SK(+) |
EcoRI |
PCR |
GGAATTCAAGGAAGTTCAGAAGCAGCAAG |
GGAATTCCCCACTCCTCATTCCTGAAATA |
n.a. |
in situ
hybridization |
|
40 |
aug3.g8337 |
LOC107456131 |
pTriplEx2 |
SfiI |
Pt-gata5 / EST clone
(LC671430/eS6_d1_049_F06) [ref. 2] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
41 |
aug3.g4322 |
LOC107451809 |
pZL1 |
SalI/NotI |
Pt-hedgehog (hh)/pZL-km-hh#10-2 (AB125742) [ref. 4] |
n.a. |
n.a. |
n.a. |
in situ
hybridization; |
|
42 |
aug3.g4322 |
LOC107451809 |
pBluescript II
SK(+) |
EcoRV |
Pt-hedgehog (hh)/pBS-km-hh#10-2_SalSpe#5 (AB125742) [ref. 4; denoted as At-hh1] |
n.a. |
n.a. |
T7KS - T7SK |
pRNAi (ATAC-seq) |
|
43 |
aug3.g6985 |
LOC107455300 |
pBluescript II
SK(+) |
NotI |
Pt-forkhead (fkh)/pBS-km-fkh#1-2_BglXba (AB096073) [ref. 5] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
44 |
aug3.g9172 |
LOC107457189 |
pZL1 |
SalI/NotI |
Pt-orthodenticle (otd)/pZL-km-otd#8-1 (AB096074) [ref. 5] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
45 |
aug3.g15189 |
LOC107447678 |
pZL1 |
SalI/NotI |
Pt-msx1/At_eW_020_B15 (LC379630) [ref. 6] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
46 |
aug3.g25248 |
LOC107456525 |
pZL1 |
SalI/NotI |
Pt-Delta/pZL-km-Delta-dT#10-1 (AB287420) [ref. 3] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
47 |
aug3.g16926 |
LOC107442925 |
pBluescript II
SK(+) |
EcoRI |
Pt-decapentaplegic (dpp)/pBS-km-dpp_NcoEco (AB096072) [ref. 5] |
n.a. |
n.a. |
n.a. |
in situ
hybridization |
|
48 |
non-spider gene |
n.a. |
pBluescript II
SK(+) |
EcoRI |
green fluorescent protein
(from pQBI-25 [Quantum Biotechnologies]) |
n.a. |
n.a. |
T7KS - T7SK |
pRNAi
(specificity validation; phenotype analyses) |
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*Sequences of
primers indicated by short names are as follows: |
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T7KS,
TAATACGACTCACTATAGGGAGACCACTCGAGGTCGACGGTATC |
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T7SK,
TAATACGACTCACTATAGGGAGACCACCGCTCTAGAACTAGTGGATC |
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T7new,
TAATACGACTCACTATAGGG |
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T7_5TrpEXLD2,
TAATACGACTCACTATAGGGCTCGGGAAGCGCGCCATT |
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[ref. 1]
Yamazaki et al. Zoolog. Sci. 22: 177-185 (2005) |
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[ref. 2]
Kanayama et al. Nat. Commun. 2:500 (2011) |
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[ref. 3] Oda et
al. Development 134:
2195-2205 (2007) |
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[ref. 4]
Akiyama-Oda and Oda, Development 137: 1263-1273 (2010) |
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[ref. 5]
Akiyama-Oda and Oda, Development 130: 1735-1747 (2003) |
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[ref. 6]
Akiyama-Oda and Oda, Sci. Adv. 6: eaba7261 (2020) |
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