Handbook of Meat Processing

506 Chapter 29

GMO Target sequence Technique Reference

Rapeseed Falcon6/Ac,

HCN10, Liberator6/

Ac, HCN28, HCN92

EPSPS, pat , P - 35S y PEPC RTi - PCR Zeitler et al., 2002

Potato B33 npt II, gbss - as; B33, T - DNA cPCR Hassan - Hauser et al.,

1998

Potato B33 - INV aphIV cPCR Anonymous et al.,

1997

Potato NewLeaf Plus PLRV - rep gene cPCR Hernandez et al.,

2004b

Potato NL Russet

Burbank and NL

Superior

P - 35S and cry 3A cPCR Mel ’ nychuk et al.,

2002

Tomato FlavrSavr ™ npt II, d - P - 35S/

polygalacturonase

cPCR Meyer 1995a, b

Tomato Zeneca T - nos , polygalacturonase cPCR Busch et al., 1999

Maize Bt176, MON810,

Bt11, GA21 and T25)

and soybean Roundup

Ready

Different junctions RTi - PCR fi ve GMO Kuribara et al., 2002

Maize Bt176, MON810,

Bt11, GA21 and

soybean Roundup

Ready

Cp4EPSP/NOS terminator,

35SCamV/BAR, 35ScaMV/

adh, 35ScaMV/hsp70, actin

promoter/CTP

RTi - PCR fi ve GMO Peano et al., 2005

Maize Bt176, Bt11,

GA21, and soybean

GT73

Different junctions RTi - PCR four

GMO

Taverniers et al.,

2005

Maize Bt176, MON810,

Bt11, and soybean

Roundup Ready

Different junctions RTi - PCR four

GMO

Greiner et al., 2005

Maize Roundup ready

and soybean Roundup

ready

CTP/EPSPS RTi - PCR Lerat et al., 2005

Table 29.1. GMO specifi c PCR methods for GMO analysis. cPCR: conventional PCR; RTi-PCR: real-
time PCR; dc-PCR: double competitive PCR; qc-PCR: quantitative competitive PCR. Modifi ed from
Hernandez et al. (2005) (cont.)

ated: a “ nonspecifi c strategy ” independent
of the target sequence (e.g., through fl uores-
cent dyes that have special fl uorescent prop-
erties when bound to double stranded (ds)
DNA), and a “ sequence - specifi c strategy ” by
sequence - specifi c fl uorescent oligonucle-
otide probes. The latter strategy also allows
the simultaneous detection of multiple
targets, using multiple probes labeled with
different reporter dyes (Bernard et al. 1998 ;
Wittwer et al. 2001 ; Dupont et al. 2002 ). This
is of special interest in GMO analysis, since

a wide diversity of transgenic events is intro-

duced in the market.

Several fl uorescent dsDNA - specifi c dyes

have been used in RTi - PCR methods. The fi rst

used was ethidium bromide (Higuchi et al.

1993 ; Le Pecq and Paoletti 1996 ; Wittwer

et al. 1997 ), and subsequently, other interca-

lating dyes have also been introduced, such as

YO - PRO - 1 (Ishiguro et al. 1995 ; Tseng et al.

1997 ) and SYBR Green I (Hernandez et al.

2003c ). They are cost - effective, as it is not

necessary to add a target - specifi c fl uorescent