Journal: BMC Biotechnology

Article Title: GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use

doi: 10.1186/1472-6750-12-15

Figure Lengend Snippet: Estimation of genomic DNA quantity and quality . Bar graph represents average estimated DNA concentrations from at least 50 extracted samples ( A ; ng/μl), as defined by gel density analysis (white bars), or spectrophotometric determinations using a NanoDrop spectrophotometer (blue bars). Results were obtained for DNA samples extracted using Promega's Genome Wizard kit ( I ), the Nucleon Phytopue Kit ( II ) or the CTAB-homebrew method ( III ). Collated absorbance ratios obtained from the same samples at 260: 280 nm (green bars), or 260: 230 nm (grey bars) are shown. DNA quantity and integrity was assessed by resolving DNA samples (extracted from Bt11 maize reference material; 0%, - 5%) on 0.8% TAE agarose gels by electrophoresis. Representative samples extracted using Promega's Genome Wizard kit ( B ), the Nucleon Phytopue Kit ( C ) or the CTAB-homebrew method ( D ).

Article Snippet: The quality of maize genomic DNA samples, extracted from Bt11 seed reference material were compared after extraction, using three different and commonly used plant DNA extraction methods (CTAB; Nucleon PhytopureTM; Promega Genome WizardTM).

Techniques: Spectrophotometry, Electrophoresis

Journal: BMC Biotechnology

Article Title: GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use

doi: 10.1186/1472-6750-12-15

Figure Lengend Snippet: Influence of the DNA extraction method on RT-PCR and LAMP-BART efficiency . LAMP-BART (red points) and RT-PCR (black points) standard curves made by serial dilutions of DNA isolated using Promega's Genome Wizard kit ( A ), the Nucleon Phytopure Kit ( B ) or the CTAB-homebrew method ( C; diluted 10 fold to reduce inhibitory nature of this preparation). DNA isolated using each extraction technique was amplified with the CaMV 35 S-p (1), NOS-t (2) or ADH1 (3) primer sets. Template concentration in LAMP-BART comparisons are plotted as a function of time to light peak (T max ), while RT-PCR comparisons are based on cycle number (Ct). Each RT-PCR cycle is approximately equivalent to 2 min.

Article Snippet: The quality of maize genomic DNA samples, extracted from Bt11 seed reference material were compared after extraction, using three different and commonly used plant DNA extraction methods (CTAB; Nucleon PhytopureTM; Promega Genome WizardTM).

Techniques: DNA Extraction, Reverse Transcription Polymerase Chain Reaction, Isolation, Extraction, Amplification, Concentration Assay

Journal: BMC Biotechnology

Article Title: GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use

doi: 10.1186/1472-6750-12-15

Figure Lengend Snippet: Regression analysis was performed on PCR and LAMP-BART data sets obtained using various DNA extractions performed on a titration series of Bt11 maize reference tissue (0.1 - 5%; nd - not determined)

Article Snippet: The quality of maize genomic DNA samples, extracted from Bt11 seed reference material were compared after extraction, using three different and commonly used plant DNA extraction methods (CTAB; Nucleon PhytopureTM; Promega Genome WizardTM).

Techniques: Titration, Amplification, Extraction, Sequencing

Journal: BMC Biotechnology

Article Title: GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use

doi: 10.1186/1472-6750-12-15

Figure Lengend Snippet: Comparison of inhibitors of PCR and LAMP-BART . Bt11 maize genomic DNA samples (5% GM) were subjected to RT-PCR amplification using ADH1 primers (A); CaMV 35 S-p primers (C) or NOS-t primers (E). A comparative set of analysis was performed using LAMP-BART, conducted in the presence of ADH1 primers (B); CaMV 35 S-p primers (D) or NOS-t primers (F). The extracted sample contained either 10 5 copies of wild type genome or 5 × 10 3 copies of transgenic genome. The analysis was performed in the presence and absence of final inhibitor concentration defined: sodium dodecyl sulphate; (SDS; 0.005% [1]; 0.01% [2] – GREEN BARS); cetyl trimethylamonium bromide (CTAB; 0.005% [1]; 0.01% [2] – BLUE BARS); sodium chloride (NaCl; 25mM [1]; 50mM [2]- BLACK BARS); xylan (XL; 0.1% [1]; 0.25% [2] – RED BARS); rice starch (RS; 0.1% [1]; 0.25% [2] – GREY BARS); humic acid (HA; 0.01ng [1]; 0.1ng [2] – BROWN BARS); calcium chloride (CaCl 2 ; (1µM [1]; 100µM [2] – WHITE BARS). The broken line (----------), represents the average Ct or Tmax value for each set of amplifications performed in the absence of inhibitor. In some instances amplification or reporting of this activity was completely inhibited (not determined; Nd ).

Article Snippet: The quality of maize genomic DNA samples, extracted from Bt11 seed reference material were compared after extraction, using three different and commonly used plant DNA extraction methods (CTAB; Nucleon PhytopureTM; Promega Genome WizardTM).

Techniques: Comparison, Reverse Transcription Polymerase Chain Reaction, Amplification, Transgenic Assay, Concentration Assay, Starch, Activity Assay