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Table 2 Details of primers designed with AlignMiner to identify specifically by PCR the five A. thaliana GS1 genes as well as the two primer pairs that identify the photosynthetic and non-photosynthetic isoforms of P. pinaster; note that the 3' (reverse) primer is complementary to the sequence appearing in Figures 6 and 8.

From: AlignMiner: a Web-based tool for detection of divergent regions in multiple sequence alignments of conserved sequences

Isoform

Primer

Length

%GC

Tm (°C)

Amplicon size (bp)

GS1.1

5'-GGTCTTTAGCAACCCTGA-3'

18

50

54.6

740

 

5'-ATCATCAAGGATTCCAGA-3'

18

39

48.7

 

GS1.2

5'-GATCTTTGCTAACCCTGA-3'

18

44

51.3

739

 

5'-CTTTCAAGGGTTCCAGAG-3'

18

50

53.6

 

GS1.3

5'-AATCTTCGATCATCCCAA-3'

18

39

50

739

 

5'-AAAGTCTAAAGCTTAGAG-3'

18

33

46

 

GS1.4

5'-GATCTTCAGCCACCCCGA-3'

18

61

59.4

739

 

5'-AATGTGTCATCAACCGAG-3'

18

44

51.5

 

GS1.5

5'-GATCTTTGAAGACCCTAG-3'

18

44

48.8

740

 

5'-TCTTTCATGGTTTCCAAA-3'

18

33

50.1

 

Photosyntetic isoform

5'-AGTGCGCATTAAGGACCCATCA-3'

22

50

61

177

 

5'-ACACACTGGCTTCCACAATAGG-3'

22

50

59.4

 

Non-photosynthetic isoform

5'-ACAGATGATCTAGGACATGC-3'

20

45

52

169

 

5'-CACTTATTTGCACTTGAAGG-3'

20

40

52.6