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. 2014 Apr 22;9(4):e95877. doi: 10.1371/journal.pone.0095877

Table 1. Primers used to isolate/analyze GL1, GL2, GL3, TTG1 and TRY genes from B. villosa.

Primers NCBI accession Sequence (5′ to 3′)
Coding Sequences from B. napus 1
GL1-F HQ162473.1 GTCAGGATCCATGAGAACGAGGAGAAGAACAGA
GL1-R GTCACTGCAGCTAGAGACAGTAGCCAGTATCA
GL2-F EU826520.1 GTCAGGATCCATGTCAATGGCCGTCGAGATGTCA
GL2-R ATATCTGCAGTGTTGTGCAGCGTGACAGAGACGA
TRY-F EE451172.1 AGCTGGATCCGCTTGCATTCTCCAACT
TRY-R CGCACTGCAGGCAATTTCGTTATGCTATATG
Coding Sequences from B. rapa 1
EGL3-F HM208589.1 GTCAGGATCCATGGCTGCTGTAGAAAACAG
EGL3-R GCAGCTGCAGAGTGCATCTTGAATCATTCCT
TTG1-F HM208590.1 TAGAGGATCCATGGACAACGCAGCTCCGGACT
TTG1-R AGTCGGTACCTCAAACTCTAAGGAGCTGCA
Conserved Regions (for Q-PCR) 2
GL1-F ACTGGGCTGAAGAGGTGTGGA
GL1-R GAGATGAGTGTTCCAGTGA
GL2-F CGCTGGCCGGGAGAAAGAGC
GL2-R GGAGGTTTTTTCTGGATGAA
EGL3-F ACATTCAATGGAGTTACGGA
EGL3-R AGAGATTCGTAAAGCTCTCT
TTG1-F CTCTGGGAGGTCAACGAA
TTG1-R ATGCTGCACGTGCCTAAC
TRY-F CATCACTCCTCTTCTCACA
TRY-R TGTGGTGGGGAAGAAAACAGA
BnEF1F (B. napus) CCCATTCGTCCCCATCTCTGGA
BnEF1R (B. napus) ACGGAGGGGCTTGTCCGAGG
1

Forward (F) primers were designed at the 5′ coding end and reverse (R) primers to the 3′ end of cDNA sequences from B. napus for GL1 and GL2 and from B. rapa for EGL3 and TTG1. Forward primers for TRY were designed 72 nucleotides before coding region and reverse primer at 3′ end of the cDNA from B. rapa. Underlined sequences indicate incorporated restriction enzyme sites; BamHI and PstI sites in the GL1, GL2, EGL3 and TRY forward and reverse primers, respectively; BamHI and KpnI sites in the TTG1 forward and reverse primers, respectively.

2

Primers for Q-PCR were designed to conserved regions based on alignments between the A. thaliana homologue and the homologues from four Brassica species. B nEF1F/BnEF1R are endogenous reference gene primers.