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Table 4 Primers and fluorogenic probes for Real-time PCR

From: Critical points of DNA quantification by real-time PCR – effects of DNA extraction method and sample matrix on quantification of genetically modified organisms

Target/amplicon length

Specificity/sequence accession number

Primer and probe names

Primer/probe sequence

Lengh (bp)

Reference

Lectin

81 bp

soybean specific gene (Accession number K00821)

TM-Lectin-F

5'-tcc acc ccc atc cac att t-3'

19

29

  

TM-Lectin-R

5'-ggc ata gaa ggt gaa gtt gaa gga-3'

24

 
  

Lectin-FAM

5'-FAM-aac cgg tag cgt tgc cag ctt cg-TAMRA-3'

23

 

Invertase

79 bp

maize specific gene (Accession number U16123)

ivr1-TM1

5'-tgg cgg acg acg act tgt-3'

18

30

  

ivr1-TM2

5'-aaa gtt tgg agg ctg ccg t-3'

19

 
  

ivr

5'-VIC-cga gca gac cgc cgt gta ctt cta cc-TAMRA-3'

26

 

p35S

82 bp

p35S promoter (Accession number V00141)

TM-35S-1

5'-gcc tct gcc gac agt ggt-3'

18

29, 31

  

TM-35S-2

5'-aag acg tgg ttg gaa cgt ctt c-3'

22

 
  

35S

5'-FAM-caa aga tgg acc ccc acc cac g-TAMRA-3'

22

 

tNOS

151 bp

tNOS terminator (Accession number V00087)

tNOSF

5'-gtc ttg cga tga tta tca tat aat ttc tg-3'

29

32

  

tNOSR

5'-cgc tat att ttg ttt tct atc gcg t-3'

25

 
   

5'-FAM aga tgg gtt ttt atg att aga gtc ccg caa-TAMRA-

  
  

tNOSpro

3'

30

 

GTS 40-3-2 soybean

83 bp

construct specific (Accession number AB209952)

RRS-F

5'-gcc atg ttg tta att tgt gcc at-3'

23

29

  

RRS-R

5'-gaa gtt cat ttc att tgg aga gga c-3'

25

 
   

5'-FAM-ctt gaa aga tct gct aga gtc agc ttg tca gcg-

  
  

RRS

TAMRA-3'

33

 

MON810 maize

115 bp

event specific (Accession number AF434709)

Mon810F1311

5'-cct tca taa cct tcg ccc g-3'

19

33

  

Mon810R1311

5'-aat aaa gtg aca gat agc tgg gca-3'

24

 
   

5'-FAM-acg aag gac tct aac gtt taa cat cct ttg cca-

  
  

Mon810pro1311

TAMRA-3'

33