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Table 3 Sensitivity analysis of the bidirectional DNA walking method, including the Cry-F and Cry-R walking direction, using the four different mixes of DRT primers (A-D)

From: Development and validation of an integrated DNA walking strategy to detect GMO expressing cry genes

 

Cry-F

 

2000 HGEs

200 HGEs

20 HGEs

Bt11 maize

+

+

+

A

B

C

D

A

B

C

D

A

B

C

D

550

500

650

950

1250

450

1000

1400*

1800

550

500

 

450

1000

1400*

1800

550

  

1000*

1400

1800

MON810 maize

+

+

+

A

B

C

D

A

B

C

D

A

B

C

D

550*

850

1300

650

 

400

   

400*

  

1400

400*

T304–40 cotton

+

+

+

A

B

C

D

A

B

C

D

A

B

C

D

550

800

350

450

500

600

800*

1200

400

600

550*

800

1000

500

1000

450

500

600

800

1200

400

500

1000

1700

250

550*

500

 

400

Cry-R

Bt11 maize

+

+

+

A

B

C

D

A

B

C

D

A

B

C

D

400*

800

> 2000

500

1000

1400

550

1300

1600

300

400*

> 2000

1200

300

550

1700

300

400*

 

500

1000

550

1000

MON810 maize

+

+

+

A

B

C

D

A

B

C

D

A

B

C

D

250*

770

350

500

260

250*

600

500

350

250*

350

 

350

T304–40 cotton

+

+

(+)

A

B

C

D

A

B

C

D

A

B

C

D

550

200

200

450

550

200

450

700*

250

550

200*

1000

550

250

450

250

200*

550

450

  1. For each tested sample (Bt11, MON810 and T304–40), 2000, 200, 20 HGEs of the target were tested. For each result, the experiment was carried out in triplicate. The detection of the targets is symbolized by + (amplicons observed in each repetition) or (+) (amplicons not observed in each repetition). The approximate size of amplicons is indicated in base-pair for each DRT primers. The amplicons observed in each repetition (3/3) are indicated in bold while the amplicons obtained not in each repetition (1–2/3) are indicated in italic. The sequenced amplicons are indicated by an asterisk