Table 2.
Primers used for qPCR and generation of in situ RNA hybridization probes
| Gene (SPU_#) | Primers for qPCR | Primers for in situ probe | RNA in situ probe gene location |
|---|---|---|---|
| Sp-Pho (10295) | F: TGTCAACCCATCTCGATTCA R: CAGGGACAGAGTCTGCATCA |
F: GAGGTGGAGATTGAGAGCATACCAATGG R: AAAGGCAACGTGACTACCACTGTCTG |
1100 nucleotides, includes 4 C2H2 Zn-finger domains |
| Sp-Mbt1 (13689) | F: ACTCGATGGATGGCAGTACC R: TACACCAGGGTGGGAAGAAG |
F: CAGATGTCGGAGGAAGATGAACTGAAGC R: TAGGAATTGAACCACCTCAGCGATTCC |
1104 nucleotides, includes C-terminus |
| Sp-Mbt2 (21547) | F: CTGATCAACGCTGCCTACAA R: ATCATGGCACGGTAGCTCTT |
F: GCTTCTCTGGTCCTTACCTGAACAAGG R: TGAATACTTTGGCAAGACTGGCACAGTC |
1102 nucleotides, includes C-terminus |
| Sp-L(3)mbt3 (17786)➔(21123)* | F: TCACTTTGATGGTTGGGACA R: GCCTGATTGCCCAGTGTTAT |
F: CAGTTGGACTGCTTACCTAAAGATGACCAG R: GGACGTCTTATGATGACTGGTGTACTTCC |
922 nucleotides spanning both gene predictions, includes 2 MBT and 2 C2H2 Zn-finger domains |
| Sp-Pc1 (20586) | F: TCGCTCCGAACTTATCCCTA R: GTTTTCGTTCTTCGCTTTCG |
F: TTATAGATGAGAAACCGTTGAGCAAACCCG R: TCAGATCCAGTGGCGAATCCACC |
1075 nucleotides, includes C-terminus |
| Sp-Pc2 (20946) | F: AACAATGAGGATGGGGATGA R: TCAACATCGACATTGGGAGA |
F: CTCTGGGAAGACCTCCGTTGTTTCG R: CTACGAGGCGGACTTTATAAGTGATGACG |
1064 nucleotides, includes C-terminus |
| Sp-Ph (00345) | F: GCTCCAAGAGACTGGGTCTG R: TCCAAGGTTGTGCCACATTA |
F: CAGCTCTACAACCATTCCAACACAGTCC R: GCTCAAGGCTGTCATCAAATGATCCTCC |
1042 nucleotides, includes 70% of SAM domain |
| Sp-Psc (23424) | F: AGTAGCTGCGGTCCGATAAA R: CAGCTGCAGGACATCTCAAA |
F: CGCAAAACGCGGCTGAAGATAACG R: GGGCATTGTAGGTATGCTCAACGC |
1404 nucleotides, includes C3CH4 Zn-finger domain |
| Sp-Ezh2 (23366) | F: TGCTACCTCGCTGTGAGAGA R: TTTCCATCCTCGTTGGAGAC |
F: ACTGTTCTTCACAACATTCCTTACATGGGC R: TGGCATGTTGGAATGATTGGCAAAACG |
1587 nucleotides, includes SET domain |
| Sp-Eed (19607)➔(26082)* | F: CCCCTACAAGCATGGTGACT R: CTTCCGATACGGGTTGAAGA |
F: ATAAACAAAGCGGGTGTTTCAAACGTCGG R: CTTGATCATCTCTACAGGGTCATGAATGCC |
1083 nucleotides spanning both gene predictions, includes 4 WD domain repeats |
| Sp-Suz12 (13163) | F: TTCCAGAGGAATGGACCTTG R: GGGTGCGGAAATACGTTCTA |
F: ATGGCTCCCCACAAACATTCTCGG R: TCCATTCCTCTGGAAGGCATAGCC |
1509 nucleotides, includes N-terminus |
| Sp-Rbbp4 (26263) | F: ATCTTGCCAAAGCGAGTCAT R: CAGATCCCACAATGCAACAG |
F: ATGAGCAGAACCACCTAGTCATAGCCAG R: CAGTCTGTTGTTGGCATTCTAAATCAGCC |
1062 nucleotides, includes 5 WD domain repeats |
*
These genes each consist of two overlapping gene predictions
Ubq F: CACAGGCAAGACCATCACAC
Ubq R: GAGAGAGTGCGACCATCCTC
Primers are written in the 5′ to 3′ direction