It was recently suggested that it is preferable to expose gDNA to endonuclease restriction in order to improve amplification efficiency and to increase the accuracy of GMO target copy number measurement with cdPCR [19]. Dairy products: the change in consumption over the past decade. Upper frame: Non digested DNA. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. A limited specificity study was however conducted on two samples containing either wild-type maize, or DNA from milk sample without maize. To sum up, this collection of food-related research topics is quite extensive. Both qualitative and quantitative research falls under consumer research and is normally completed by market research companies that specialize in this type of research. These fast food research questions are as relevant as never. However, most of the proposed solutions are not practical and reliable quantification of GMOs in food and feed samples still remains a major challenge. Acceptance criterion for trueness is that the measured value has to be within ±25% of the accepted reference value over the whole dynamic range [24], [25]. Based on the experience acquired during this study and assuming the samples and mixes are already prepared, the simultaneous analysis of these four samples with ddPCR would require approximately 190 minutes. Quantitative Research Titles about Cookery & Food, Qualitative Research Titles about Cookery & Food, 204 Research Topics on Technology & Computer Science, 497 Interesting History Topics to Research, Making school lunches healthier and more affordable, Healthy food: the impact of the vegetarian diet, Nutrition and food security within the aboriginal and remote communities of Australia, Food safety: a policy issue in agriculture today, Staff turnover in restaurants: causes and effects, 280 Good Nursing Research Topics & Questions, 226 Research Topics on Criminal Justice & Criminology, 180 Best Education Research Topics & Ideas, 110+ Micro- & Macroeconomics Research Topics, 417 Business Research Topics for ABM Students, 190+ Research Topics on Psychology & Communication, 281 Best Health & Medical Research Topics, 501 Research Questions & Titles about Science, A List of Research Topics for Students. Customized Food Research Paper Topics for College Students. Is the atmosphere that important for restaurants? The results obtained should be applicable to other fields where quantitative testing in food and feed samples is required. NFO DONOVAN RESEARCH – Food Labelling Issues - Quantitative Research with Consumers C02020 TABLE OF CONTENTS EXECUTIVE SUMMARY 1 NEEDS AND ATTITUDES TOWARDS FOOD ... November 2000 (gazetted in December 2000) and was implemented by the food industry over a two-year transition period, ending in December 20021. Time needed for quantification with qPCR and ddPCR. Top 100 Research Topics & Titles about Food & TVL When you look for a good research paper topic, you can easily become the severest critic of any proposed idea. A quantitative approach is all about collecting and analyzing numerical data. Increasing the multiplexing will certainly give further, additional advantage to ddPCR in terms of cost and throughput, and could allow its use already at the screening and/or identification steps. 5 - GMO labeling: Right to know or bizarre new precedent? Salmonella cases in port cities: a quantitative evaluation. For more information about PLOS Subject Areas, click No, PLOS is a nonprofit 501(c)(3) corporation, #C2354500, based in San Francisco, California, US, https://doi.org/10.1371/journal.pone.0062583, http://gmo-crl.jrc.ec.europa.eu/summaries/Mon810_validation_report.pdf, http://www.codexalimentarius.org/download/standards/11667/CXG_074e.pdf, http://gmo-crl.jrc.ec.europa.eu/doc/Min_Perf_Requirements_Analytical_methods.pdf. If you think about it carefully, most of our lives are spinning around food. Both stock and diluted DNA solutions from the CTAB extract could be used for MON810 content determination with qPCR. No, Is the Subject Area "Genetically modified organisms" applicable to this article? Everything from agriculture and food processing to distribution and finances is included in the definition of the food industry. This book focuses on the food safety challenges in the vegetable industry from primary production to consumption. It is noteworthy that the deviation between the MON810 content measured by ddPCR and the reference value tended to increase with lower target copy number. aLOD was estimated to be five copies for the hmg system, and six copies for the MON810 system, which is suitable for routine GMO testing. Results from the dilution series used for the dynamic range, the aLOD and aLOQ determination, and the overall repeatability. GMO content in food and feed samples is expressed in relative terms as the ratio of the quantity of the transgene (GM target, i.e. This was further confirmed by the successful establishment of several additional duplex ddPCR assays from singleplex qPCR assays, targeting GM events or screening elements, without optimization (data not shown). In the wheat sample, the generally used NucleoSpin Food kit (NSF) extraction protocol resulted in strong inhibition of the qPCR amplifications, and new DNA extraction following the CTAB protocol was needed. GMO laboratories use adapted DNA extraction methods that in some cases may reduce inhibition during the qPCR analysis. The ddPCR system is shown to offer precise absolute and relative quantification of targets, without the need for calibration curves. Digestion of DNA may be necessary for digital PCR analysis in the case of a possible linkage between targets, such as multiple copies of target physically linked on the same chromosome, or if different targets present on a same plasmid need to be quantified. By comparison, the MON810 content measured by qPCR was close to the limit of acceptance (Table 1). Lower frame: TaqI digested DNA. Upper right quadrant: FAM (hmg) positive-VIC (MON810) positive droplet cluster. Students can find guides on how to improve their writing and academic performance in general, get useful tips on making their study process more efficient, or learn how to deal with particular issues. False-positive rates observed with ddPCR. Inter- and intra-cartridge repeatability. At a higher concentration (118,000 hmg copies per 20 µl ddPCR mix), each droplet contained, on average, 5.9 hmg molecules, which is the upper recommended concentration for use of the droplet system (Bio-Rad, personal communication). In the same research determine th. Table S8. This sample questionnaire template consists of 20 questions for food quality evaluation, customer service, the value of food, hygiene levels, and overall customer satisfaction. Another difficulty linked to GMO detection is that the event-specific targets needed for reliable and specific GMO quantification are usually the junction regions between the transgene and the host plant genome, leaving a very narrow window for design, and decreasing further the flexibility for a multiplex design. A recent study has estimated that the theoretical ddPCR dynamic range is 105 target copies, and it has been established experimentally that the dynamic range covers more than 4 orders of magnitude [15]. The absolute limit of detection (aLOD) is the lowest target copy number in a sample that can be reliably detected, but not necessarily quantified [25]. The long-term effects of consuming fast food on health. This characteristic has the advantages of reducing the cost of analysis, and of decreasing the uncertainty linked to droplet volume variation and dilution pipetting errors [19]. Therefore, a research title about a restaurant would be a perfect choice for someone who wants a career in this area. Quantitative, as the name suggests, is about surveying your market to gain a large quantity of information which can be processed numerically, then data-modelled enabling you to identify trends, preferences and attitudes. The basis of digital PCR (dPCR) is to quantify the absolute number of targets present in a sample, using limiting dilutions, PCR and Poisson statistics [14]. What are the advantages of the macrobiotic diet? However, the hmg and MON810 were affected differently under all the tested conditions, resulting in under-estimations of the MON810 content and/or loss of sensitivity (data not shown). Comparison of duplex ddPCR quantification on digested and non-digested genomic DNA. Ask our academic expert for help with your assignment! In the absence of a DNA reference material certified for absolute copy number concentration, trueness could only be assessed for the MON810 content. No, Is the Subject Area "Maize" applicable to this article? The aLOQ of the duplex ddPCR assay was therefore equal to or better than that estimated for the qPCR performance. One of the limitations is the need to choose target sequences with similar, short lengths. Table S12. Then, we moved on to the most relevant topics in our fast-paced world: fast-food and restaurants. A total of 35 replicate positive ddPCR results were then analyzed. Upper left quadrant: FAM (hmg) positive-VIC (MON810) negative droplet cluster. https://doi.org/10.1371/journal.pone.0062583.s004. People hate waiting for their food for too long, so the fast-food demand is growing. And, of course, the top 10 food research questions for your paper! ?” That’s not what I mean. Several advantages are proposed for the use of dPCR instead of qPCR in routine GMO testing: 1) it enables absolute target copy number to be detected and avoids the amplification efficiency bias observed with qPCR [11], [19], 2) it overcomes dependence on the availability of references or standards [18], 3) it provides data with the high precision and confidence necessary for metrological use [11], [19], 4) it provides more accurate data at low target copy numbers than qPCR [20], allowing quantification of low GMO content, and 5), because of its tolerance to inhibitors as an end-point measurement, it can reduce the biases linked to matrix type often observed with qPCR [18]. How to achieve the umami flavor in your cooking? the nucleic acid fragment introduced in the host genome) to that of the endogene (reference gene in the host genome) [10]. The food and restaurant market is a vast and demanding industry that is expanding at a rapid pace. Funding: The work was supported by the Slovenian Research Agency (grant Nos. To be employed in routine testing, methods based on ddPCR shall be properly validated through ring-trials and verified during their introduction in laboratories to demonstrate their fitness for the purpose. Canning and preserving meat: a case study. Linearity over the dynamic range was determined by the coefficient of correlation R2, calculated on the average of the target copy numbers measured in the replicated dilution series for both qPCR and ddPCR. How to eat your eggs: royale vs. Florentine? Currently, the most common technique for analyzing the presence of nucleic acids in food and feed samples is the polymerase chain reaction (PCR) [1]–[3]. An attempt to optimize the duplex qPCR assay was made by varying primers and probe concentration of both systems. Considering the cost of reagents, consumables and labor at NIB and the above proposed set-ups for GMO testing, the quantification of a given transgene in four samples with ddPCR would cost approximately US$20.9 per sample and US$22.3 using qPCR. Acceptance criterion for repeatability is ±25% of the target content (from 0.58% to 0.96%) represented by the dashed lines. 6 µL of digested gDNA were analyzed on a 1% agarose gel to confirm complete digestion. Chemicals of Potential Concern Less than 10% variability was observed within each of the five cartridges for the determination of hmg copies, MON810 copies, and MON810 content. What are the best oils for frying: saturated fat issue? Discover a faster, simpler path to publishing in a high-quality journal. Recently, a cdPCR commercial system has been demonstrated that enables suitable metrological performance for the certification of the copy number ratio of reference materials used in GMO testing [11], [19]. VIC vs. FAM channel clustering plot of droplets for non-digested and TaqI digested MON810 DNA. Throughout the dynamic range, each individual ddPCR measurement of the MON810 content fell within ±25% of the certified value (Figure 3). Custom-writing.org team created this list of research titles. The intention of this study was to demonstrate the usability of ddPCR in real-life routine diagnostics, rather than to re-investigate the recently reported metrological characteristics of this technology [15]. All samples were stored at −20°C. If you are not sure how to choose a food research topic, here is a short guide for you. However, private farming is not considered to be part of the food industry. MON810 contents measured by the ddPCR duplex MON810/hmg assay in maize seed-powder flour samples and corn flakes samples are in good accordance with the values measured with the qPCR singleplex MON810 and hmg assays (Table 1). Droplets were generated for each individual cartridge, and those droplets containing the PCR mixes of the three cartridges were transferred onto a single PCR plate for amplification followed by droplet count. gDNA endonuclease restriction for ddPCR testing was also evaluated and found unnecessary (Table S4). Healthy fast-food chains in the US: an overview. Some of them are also certified for the copy/copy (cp/cp) transgene/endogene ratio (see Appendix S1 and Table S1). Food and cookery is a wide area, which means it includes all the issues related to food. Whether it is the changing dynamics of dining or the latest in chain restaurant geo-location, food industry market research requires the most advanced, up to date information to create successful strategy. https://doi.org/10.1371/journal.pone.0062583.s007. Quantitative outcome research is mostly conducted in the social sciences using the statistical methods used above to collect quantitative data from the research study. Quantitative research is most common encountered as part of formal or conclusive research, but is also sometimes used when conducting exploratory research.Quantitative research techniques are part of primary research.. Quantitative research differs from qualitative research in the following ways:. Using qPCR–the common technique for GMO quantification-, standard curves for the quantities of endogene and transgene are prepared separately, using serial dilutions of DNA extracted from reference material [8]. Taking GMO testing as a concrete example, ddPCR key performance parameters, using the QX100 droplet system (Bio-Rad, Pleasanton, CA), were compared with current qPCR performance and with the recently studied performance of cdPCR [11], [18]. 6 - Go veggie: Boost your health and save the planet? Put simply, quantitative research involves gathering extensive data from the relevant sample size of the market sector or sub-sector that we’re focusing on. Low-glycemic index diet to treat diseases. Title: Systems Approach and Qualitative Methods in Food Related Research … Are beans enough to substitute meat proteins? The aim of this study was therefore to evaluate the application of ddPCR for quantitative analysis in food and feed samples. The MON810 content measured by qPCR was similar to that of the ddPCR value, the latter being slightly closer to the target value (Table 1). You can undoubtedly find a question you think is worth developing. Competing interests: The authors have declared that no competing interests exist. However, you can find research papers on this topic if you search using relevant keywords on Google Scholar. Linearity of response, absolute limits of detection and quantification, repeatability over the dynamic range of the ddPCR endogene and transgene assays were assessed. Example of quantitative research title about abm strand 1 See answer fpmagno fpmagno The ABM strand in high school can be a good area for quantitative research. The dilution series consisted of 14 solutions containing from approximately 118000 to 0.02 copies of hmg, and from 4300 to 0.0008 copies of MON810 per 20 µL, respectively). Results of quantification performed with CRM certified for transgene/endogene copy ratio were expressed as percentages of the copy/copy ratio. Similarly, the aLOQ of the duplex ddPCR assay was also in a range similar to the aLOQ measured in cdPCR (15–56 transgene copies), assessed only on the MON810 assay [18]. The absolute number of target nucleic acid molecules contained in the original sample before partitioning can be calculated directly from the ratio of positive to total partitions, using binomial Poisson statistics [15]. Also, the need is usually to quantify low concentrations of transgene (down to 0.1%) in a background of high endogene quantities. Both types can be used to start exciting research. It might save your time researching about possible topics for your research. The limits of quantification, trueness and repeatability of the duplex assay comply with international recommendations [24], [25] and are comparable or superior to those of the inter-laboratory validated qPCR singleplex assays (Table 2). The menu standards for international hotels. Self-service kiosk in fast-food restaurants. Table S10. No, Is the Subject Area "Nucleic acids" applicable to this article? An additional set of experiments was performed to establish repeatability between different emulsification runs. Fast Food Research Paper Topics for Fast Food Lovers and Haters Fast food is firmly established in our lives, and not everyone can resist the temptation to taste it. The impact of a visual representation of a dish on a customer’s review. This type of research is used by different industries and even as undergraduate research. Key performance parameters of the assay were determined. Our food market clients need more than just a flavour of one of the most complex markets on earth. In all experiments and for all three parameters (hmg and MON 810 copy number, MON810 content), the coefficient of variability measured at each point of the dynamic range was far below the 25% threshold set in international guidance documents for validation of GMO testing methods [24], [25]. Yes How to Choose an Interesting Food Topic to Research? To do so, calculations were made based on the simultaneous quantification of four samples-the average number run in parallel in middle-size GMO laboratories. For this reason, evaluation of duplex qPCR and ddPCR assays was performed and compared to the performance of singleplex assays. Affiliation Copyright: © 2013 Morisset et al. Probe and primer nucleotide sequences were the same as in the inter-laboratory validated protocol [22] but the TAMRA quencher in the probes was replaced by the Black Hole Quencher 1 (BHQ-1). The absolute limit of quantification (aLOQ) and absolute limit of detection (aLOD) for qPCR and ddPCR were determined on these experimental results. That’s why they come to Mintel. There are various topics in the general category about food processing and the historical background of some dishes. We thank Prof. Dr. Roger Pain for reviewing the manuscript. Further, ddPCR performance was evaluated using non-digested gDNA. What’s the actual research you’re conducting? Similarly, no significant variation of the measured MON810 content was observed between the singleplex and the duplex ddPCR assays (bias = 5.8%) (Table S3). Click on the link to the food safety research priorities page for more information. A good research topic will not only earn you grades but would be beneficial in your daily dietary intake and appreciation of nutrition in life. The primers and probes of both the hmg and MON810 systems were mixed in the qPCR or ddPCR volumes to final concentrations equal to those in the original singleplex assays. From these results, it was concluded that the duplex ddPCR assay performs as well as the singleplex ddPCR assays without any additional optimization. Caffeine dependency in Italy: ethnographic research. The secrets of airplane meals: what affects its taste? Set-up and number of reactions needed for simultaneous quantification of four samples with qPCR and ddPCR. The hmg gene was used as the endogenous control gene for maize. Cookies Policy. It is thus concluded that ddPCR technology can be applied for routine quantification of GMOs, or any other domain where quantitative analysis of food and feed samples is needed. Five replicates for each data point. The historical analysis: how have Asian hot pots become popular in the US? Fast food: the difference between reality and advertisement. How to Conduct Market Research for Restaurants. Set-up and number of reactions needed for quantification with qPCR and ddPCR (full 96 well-plate). Real-time quantitative polymerase chain reaction (qPCR) is currently used for quantitative molecular analysis of the presence of GMOs in products. https://doi.org/10.1371/journal.pone.0062583.s010, https://doi.org/10.1371/journal.pone.0062583.s011. What are the most harmful ingredients in fast food burgers? The aggregate represents the sum of the five series. In the case of GMO quantification, the target of the event-specific assay and the target chosen for endogene quantification are present as unique copies in the host plant genome and are not linked. No, Is the Subject Area "DNA extraction" applicable to this article? aLOD was determined as the lowest concentration level for which all five ddPCR replicates resulted in at least two positive droplets per reaction. A unique, single copy DNA integration-border region of the genomic sequence and the inserted sequence element originating from CaMV (35S promoter) were used for specific detection and quantification of the MON810 event. The ddPCR duplex assay was evaluated using three 8-well cartridges containing the singleplex hmg, the singleplex MON810 and the duplex ddPCR assay respectively. Electric vs. gas oven: the effect on baked products. Qualitative analysis of natural nutritional supplements. Error bars represent the standard deviation between the replicates for each series or in the aggregate. Food regulations: a case study of food allergens. The restaurant industry is the area related to food as much as the others. 640118/2008/67 on performing activities as holder of the national standard MIRS/NIB/FITO for the amount of substance in food of plant origin. It offers a dynamic range over four orders of magnitude, greater than that of cdPCR. In our laboratory, the aLOQ estimated for the qPCR assay on the same DNA is 18 copies. The Impossible Meat in fast food restaurants. No, Is the Subject Area "Plant genomics" applicable to this article? This result suggests that the ddPCR duplex assay is more tolerant to inhibitors found in some complex food and feed matrices than the qPCR assays, as suggested earlier [18]. The sensitivity (five target DNA copies) of the ddPCR assay compares well with those of individual qPCR assays and of the chamber digital PCR (cdPCR) approach. It has already been asserted that qPCR offers a much broader dynamic range than digital PCR [16]. Tired of searching a sample? The food industry is an enormous global machine that works non-stop to provide food to people all over the world. Another important limitation is its sensitivity to the frequent presence of inhibitors co-extracted with nucleic acid from complex matrices [8]. Yes The Jamaican influence on British cuisine. The quantitative analysis of four samples using hmg and MON810 singleplex qPCR assays requires a total of 96 reactions (see set up A, Table S9 in Appendix S2). https://doi.org/10.1371/journal.pone.0062583.s005. Qualitative research isn’t about numbers – it’s about thoughts and opinions. A quantitative survey provides data on the amounts of various foods consumed by individuals and or populations; qualitative research provides information on the kind of foods consumed, food preparation procedures, food preferences, cultural influences and attitudes towards foods. Standard curves were prepared from five serial dilutions of the copy/copy ratio certified reference material ERM-BF413gk (starting from approximately 100 ng to 1 ng DNA per reaction) and used in two replicates. Applicability of the technology has been verified on representative matrices found in routine samples, and on the range of GMO content usually found in routine samples and relevant to different international labeling requests. Food adulteration and law. 10 µL of 2× ddPCR Master Mix (Bio-Rad, Pleasanton, CA) and 1 µL of each primer (final concentration of 300 nM) and probe (final concentration of 180 nM) were mixed, and 4 µL of DNA template added. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Wrote the paper: DM MM KG JZ. The demonstrated application of ddPCR for routine quantification of GMO content in food and feed samples should act as an inducement to introduce this technology in other areas where precise analytical testing is required in food and feed samples. However, two subcategories make great research titles about the cookery strand: fast food and restaurants. Continue Reading. With ddPCR, the standard curve is not needed and a duplex MON810/hmg assay with a wide dynamic range is used. The ddPCR response was linear over concentrations ranging from an average of 5 to 118,000 hmg copies (0.02% to 99.5% positive droplets) with a coefficient of correlation (R2) of 0.9990. The data is usually gathered using more structured research instruments Four series were prepared by two operators and droplet amplification reactions were performed simultaneously on the same 96-well plate. here. Garlic and onions: Ayurveda point of view. ddPCR workflow and data analysis were performed as described (see Appendix S1) [15]. dPCR has been adopted for a number of applications, including studies of copy number variation involving allelic discrimination or imbalance, single cell gene expression, hyper-methylation, detection of low copy number nucleic acid targets (reviewed [6], [17], [18]) and of point mutations. It describes existing and innovative quantitative methods that could be applied to the vegetable industry for food safety and quality, and suggests ways in which such methods can be applied for risk assessment. chips or plate per sample), would lead to a cost per sample much greater than that required for current routine detection of GMOs in most laboratories. The impact of freezing the dough on its quality. Set-up and number of reactions needed for simultaneous quantification of 23 samples with qPCR and ddPCR. Another important factor when introducing new methods or technologies for testing GMOs in food and feed is their applicability. I have answered a very similar query here: How to write the title for a thesis or an article . This number of reactions is high, due mainly to the use of inter-laboratory validated singleplex methods, the need for monitor inhibition that requires additional dilutions, and the need for a standard curve. Next, we focused on the baking industry. Transparent windows – transparency for the customers. Concept Sort – Similar to a concept test, but involves comparing more than one product (e.g., six cups versus one cup). Yes Currently, two approaches are used in commercially available dPCR systems [15], [16]. For this study, aLOD was calculated based on experimental data obtained to determine the dynamic range. You need to understand the food industry’s categories, their size, segmentation and forecasts. Given the limitations of qPCR for the quantification of GMO in food and feed samples, especially at low target levels and in some complex matrices, the use of ddPCR in routine GMO quantification was evaluated, following the generally accepted minimum performance requirements for analytical methods [24], [25]. 4 - All-natural claims: Are they worth it? Italian culinary world: the art of matching wine to the meals. Much effort has been put into improving the performance of qPCR quantification with respect to the inhibition and matrix effects [8] and the low concentration levels of targets in routine samples [12], [13]. In the case of MON810 maize, results of the two dimensional analyses of the droplet signals in digested and undigested DNA samples (Figure S1 in Supporting Information) suggests the absence of linkage between MON810 positive droplets and hmg positive droplets, thus confirming the independence of the two targets. In the experiment performed to determine the intra- and inter-cartridge repeatability (on CRM ERM-BF413ek), the average value of the pooled ddPCR data showed good agreement with the certified value (Table 1) and, in every case, the MON810 content measured by ddPCR (within the dynamic range) was within ±25% of the certified value (Figure 2). It is also very precise at very low levels of target content. Conceived and designed the experiments: DM DS. Intra- and inter-cartridge (ddPCR 8 well chips) repeatability was determined on five independent series consisting of seven replicates of the ERM-BF413ek (approximately 100 ng and an average of 46571 hmg copies and 324 copies of MON810 per 20 µl reaction) and one NTC. Per reaction was prepared with MON810 maize event, and restaurant topics these CRMs certified... Data and then carrying out mathematical, statistical, or computing techniques food allergens ’ overview come?. 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Culinary world: fast-food and restaurants, simpler path to publishing in a high-quality journal choice someone... The experiments for dynamic range came from serial dilution of a DNA reference material has been... Culinary world: fast-food and restaurants material has also been noted [ 13.. Types can be used to determine the MON810 content with qPCR and ddPCR hate waiting their!, trueness could only be assessed for the business m/m ) GM maize/wild-type maize ratios. Prepared with MON810 maize event, and the practicability of use for routine GMO testing also... Operator on a second day and ddPCR reactions were performed ( see Appendix S1 and Table S2 ) copy/copy cp/cp...