Upper frame: Non digested DNA. Enzymatic digestion of MON810 genomic DNA (gDNA) with TaqI (New England Biolabs GmbH, Frankfurt am Main, Germany) was performed as described (see Appendix S1) [19]. In this study, the applicability of droplet digital PCR (ddPCR) for routine analysis in food and feed samples was demonstrated with the quantification of genetically modified organisms (GMOs). 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. You’d want to work out what you’re going to write about first, to be honest. This sample questionnaire template consists of 20 questions for food quality evaluation, customer service, the value of food, hygiene levels, and overall customer satisfaction. This finding supports the fact that the duplex MON810/hmg ddPCR assay can be used over a wide range of target concentrations to determine the MON810 content in a given sample, and that values around 115,000 copies constitute the upper range of quantification with ddPCR. Additionally, the repeatability of the duplex assay in measurements of the MON810 content appears slightly better that of the singleplex assays. The establishment of duplex reactions is straightforward and does not need optimization, which is also encouraging. The results obtained should be applicable to other fields where quantitative testing in food and feed samples is required. This wider range of concentrations can be attributed to the large number of partitions available for reactions in ddPCR (13,606 droplets on average in this work) compared to the number (765) available for cdPCR test. https://doi.org/10.1371/journal.pone.0062583.s010, https://doi.org/10.1371/journal.pone.0062583.s011. Yes A dilution series was prepared with MON810 maize DNA extracted from the ERM-BF413gk CRM. The types of food that can cause abdominal obesity. https://doi.org/10.1371/journal.pone.0062583.s007. Electric vs. gas oven: the effect on baked products. In this study, the applicability of droplet digital PCR (ddPCR) for routine analysis in food and feed samples was demonstrated with the quantification of genetically modified organisms (GMOs). Upper right quadrant: FAM (hmg) positive-VIC (MON810) positive droplet cluster. 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. Comparison of quantification using singleplex and duplex ddPCR assays. 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). What are the properties of CBD infused foods? The amount of fats children and adults need: statistical analysis. No, Is the Subject Area "Plant genomics" applicable to this article? 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]. How to eat your eggs: royale vs. Florentine? The lack of certified reference material has also been noted [13]. Throughout the dynamic range, each individual ddPCR measurement of the MON810 content fell within ±25% of the certified value (Figure 3). Garlic and onions: Ayurveda point of view. 6 - Go veggie: Boost your health and save the planet? The impact of the size of the plate on our level of hunger. This asymmetry in concentration renders establishment of a qPCR duplex assay targeting both the MON810 transgene and the hmg endogene even more difficult. Readers will gain increased knowledge about qualitative research and greater competency in evaluating this type of research. Dairy products: the change in consumption over the past decade. This performance was similar to those of the two singleplex qPCR assays, which was linear over the same dynamic ranges (R2 hmg = 0.9939 and R2 MON810 = 0.9958) (data not shown). Quantitative research is a method that studies numerical values. The hmg gene was used as the endogenous control gene for maize. Self-service kiosk in fast-food restaurants. Set-up and number of reactions needed for simultaneous quantification of 23 samples with qPCR and ddPCR. Including NTC and quantification control reactions in the set-up, a total of 20 reactions would be necessary to reliably quantify MON810 in four samples. 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). Caffeine in different drinks: quantitative analysis. A total of eight replicate reactions per sample were performed (see Appendix S1 for more details). Finally, we wrapped the list up with the best ten questions that can inspire you to start working on your research! It has to be noted that there is substantial empirical knowledge about the efficacy of DNA extraction methods related to sample matrix. Trueness was also evaluated on two additional CRMs and on samples from the ILC-EURL-GMFF proficiency program: ddPCR results showed better agreement with the target values than qPCR and were in accordance with the trueness acceptance criterion (Table 1). 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]. Samples representative of four different maize-containing matrices from routine GMO testing were used to test the applicability: seed-powder flour, corn flakes, wheat seed-powder flour with maize contamination, and feed containing maize (see Appendix S1 and Table S1). Less than 10% variability was observed within each of the five cartridges for the determination of hmg copies, MON810 copies, and MON810 content. 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 usage of kefir in homemade baked products. To sum up, this collection of food-related research topics is quite extensive. 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. 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]. Can’t you read my question??? Error bars represent the standard deviation between the replicates for each series or in the aggregate. 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). these title examples gives you an idea of quantitative research topic Transparent windows – transparency for the customers. Inhibition effect on MON810 quantification in qPCR and ddPCR. 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. You can choose between the approaches or move to the bakery, fast food, and restaurant topics. By comparison, the MON810 content measured by qPCR was close to the limit of acceptance (Table 1). To conduct quantitative research, methods like online survey, questionnaires, sampling methods etc. How do different types of flour affect the quality of the cake? The data provided here show that, in the context of GMO quantification, ddPCR running costs are lower than those of the standard qPCR technology, given the superior throughput, and especially when numerous samples are handled simultaneously (Table 2). Unlike qPCR, quantification by ddPCR has been found to be insensitive to the amplification inhibition present in some DNA extracts. A quantitative approach is all about collecting and analyzing numerical data. https://doi.org/10.1371/journal.pone.0062583.t001. In this research method, researchers and statisticians deploy mathematical frameworks and theories that pertain to … Here are a few other tips you can use to make sure your title will be part of the recipe for an effective research paper: Make sure your research title describes (a) the topic, (b) the method, (c) the sample, and (d) the results of your study. Error bars represent the standard deviation of the measured MON810% by ddPCR at each target concentration (five replicates per target concentration). Real-time quantitative polymerase chain reaction (qPCR) is currently used for quantitative molecular analysis of the presence of GMOs in products. There are various topics in the general category about food processing and the historical background of some dishes. The ddPCR duplex assay response was investigated over target concentrations ranging from approximately 0.02 to 118,000 hmg copies and from approximately 0.0008 to 4300 MON810 copies per 20 µL of ddPCR reaction. If you want to know more about your customers’ experiences and preferences, qualitative research is … Therefore, baking research paper topics are always relevant. broad scope, and wide readership – a perfect fit for your research every time. 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. Using ddPCR duplex assay, we have measured the absolute numbers of MON810 transgene and hmg maize reference gene copies in DNA samples. The absolute limit of quantification (aLOQ) and absolute limit of detection (aLOD) for qPCR and ddPCR were determined on these experimental results. What are the advantages of the macrobiotic diet? https://doi.org/10.1371/journal.pone.0062583.g001. Linearity of response, absolute limits of detection and quantification, repeatability over the dynamic range of the ddPCR endogene and transgene assays were assessed. The second approach, called droplet digital PCR (ddPCR), combines partitioning of the PCR test into several thousands or millions of individual droplets in a water-oil emulsion, with the use of flow cytometry to count positive PCR tests. The same primers and probes were used for both qPCR and ddPCR experiments (see Appendix S1 and Table S2). 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. Caffeine dependency in Italy: ethnographic research. Low-glycemic index diet to treat diseases. 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]. The benefits of the all-day breakfast for the business. The best ingredient for pizza Margherita: Italian chefs’ overview. However, its use for target quantification can be seriously limited by a significant bias when the target is present at low concentrations in a background of high numbers of non-target nucleic acids in the sample [4]–[7]. Dilutions of the stock extracted DNA solutions were made with nuclease- and protease-free water (Sigma-Aldrich Chemie Gmbh, Munich, Germany), using pipettes calibrated with a SAG285 precision weighing module (Mettler-Toledo d.o.o., Ljubljana, Slovenia). Results of quantification performed with CRM certified for transgene/endogene copy ratio were expressed as percentages of the copy/copy ratio. Five replicates of the dilution series and of a non template control (NTC) were measured by ddPCR. 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]. It is also very precise at very low levels of target content. Key performance parameters of the assay were determined. Insight is provided into problems that continue to plague research efforts in the food service industry. The “food halls” approach for starting businesses. 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. Research Proposal submitted to the University of Dublin Trinity College, in partial fulfilment of the requirements for the Bachelor in Nursing Studies Let’s get more specific! The target certified MON810 content (0.77%) is indicated by a dotted line. Fast food in American schools: should it be banned? we will assume that you agree to our It is so easy to make a first writing step. Ask our academic expert for help with your assignment! Consequently, diluted DNA samples were used to determine the MON810 content with qPCR. The ddPCR MON810/hmg duplex assay presented here and implemented without optimization from the inter-laboratory validated singleplex qPCR assays, achieves a wide dynamic range close to five orders of magnitude with an upper limit of quantification of about 118,000 target copies. For full functionality of this site it is necessary to enable JavaScript. To do so, the PCR mix is distributed across a large number of partitions containing zero, one or more copies of the target nucleic acid. Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia. The second phase of the study, to be published in a subsequent issue of the Journal of Hospitality Education and Research, will comprehend a considerably broader sample and seek to confirm and elaborate on these initial findings. The cookery and food industry is the largest one, among others. 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]. 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. For this study, aLOD was calculated based on experimental data obtained to determine the dynamic range. An additional set of experiments was performed to establish repeatability between different emulsification runs. One approach, termed chamber digital PCR (cdPCR), relies on the partitioning of up to a few thousand individual reactions in microfluidic chambers. Affiliation Yes Set-up and number of reactions needed for simultaneous quantification of four samples with qPCR and ddPCR. The ddPCR linear response for the MON810/hmg duplex assays covered a broader range than the same assays tested in cdPCR which was limited to 2–3 orders of magnitude [18], [19]. 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. Cookies Policy. All samples were stored at −20°C. The historical analysis: how have Asian hot pots become popular in the US? No, Is the Subject Area "Nucleic acids" applicable to this article? How do the proportions of the ingredients affect the texture of baked products? Competing interests: The authors have declared that no competing interests exist. For example, looking into only a particular country or area would make your paper more specific. Tea ceremonies in Japan as an art: ethnographic research. Browsing through these titles will give you an idea about how you can write yours. Custom-writing.org experts prepared the ultimate collection of 178 interesting food topics for research. 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). Where did the idea of steaming food come from? Conceived and designed the experiments: DM DS. 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. Nevertheless, the MON810/hmg ddPCR duplex assay met trueness acceptance criteria throughout the whole dynamic range. What are the healthiest sugar substitutes? 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. Given the larger number of replicates allowed by ddPCR than by cdPCR and the lower price per sample of the former, it has been envisaged that ddPCR could allow better precision [16], confidence and easier adoption of digital PCR technology in laboratories for daily analysis, all at lower cost per sample [15]. The impact of using plastic for preserving on the food quality. the nucleic acid fragment introduced in the host genome) to that of the endogene (reference gene in the host genome) [10]. Fast food: the difference between reality and advertisement. Table S11. The term food industry is an extremely broad one which tries to encompass an entire system that joins farms to end users through a channel of food manufacturers, wholesalers and retailers. The impact of freezing the dough on its quality. Food regulations: a case study of food allergens. The Thanksgiving dinner adaptation around the world. The ddPCR amplicons used for this study are the same as for the qPCR singleplex assays that were subjected to inter-laboratory validation and for which specificity was thoroughly checked at this stage. Before introducing a new technology in a laboratory, one has to verify its practicability for daily use [24]. 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. Table S10. The dynamic range observed for ddPCR covers the whole range of target concentrations usually needed by a laboratory for routine GMO testing (0.1% to 100% transgene/endogene ratio cp/cp). Table S9. Qualitative research isn’t about numbers – it’s about thoughts and opinions. The ddPCR system is shown to offer precise absolute and relative quantification of targets, without the need for calibration curves. Yes Everything from agriculture and food processing to distribution and finances is included in the definition of the food industry. 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. Moreover, when compared to qPCR, the ddPCR assay showed better repeatability at low target concentrations and a greater tolerance to inhibitors. qPCR efficiency and hence quantification by qPCR of endogene and transgene, can be influenced by many factors, including inhibitors, present in food and feed samples, leading to significant under- or over-estimation of GMO content [8], [11]. : P4 0165). Who is the author of this research title? What are the most harmful ingredients in fast food burgers? Then, we moved on to the most relevant topics in our fast-paced world: fast-food and restaurants. Final primer and probe concentrations (purchased at Eurofins MWG Operon, Ebersberg, Germany) in ddPCR mixes were identical to the qPCR conditions used in this study, and to those used in the previously described chamber digital PCR (cdPCR) conditions [11] (see Appendix S1 and Table S2). That is why the research design should reflect the nature of your research… Find list of quantitative research topics for American students to write good research paper. In our laboratory, the aLOQ estimated for the qPCR assay on the same DNA is 18 copies. Genetically modified food: debunking myths. 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. Finally, a limited specificity study was conducted on two samples containing either wild-type maize, or DNA from a milk sample without maize (see Appendix S1 and Table S1). https://doi.org/10.1371/journal.pone.0062583.s006. The target MON810 content (3.85%) is indicated by a dotted line. Therefore, the specificity of the ddPCR amplicons was not thoroughly verified as it is inferred that specificity should be the same as for the qPCR singleplex assays. Citation: Morisset D, Štebih D, Milavec M, Gruden K, Žel J (2013) Quantitative Analysis of Food and Feed Samples with Droplet Digital PCR. Technology trends in the restaurant industry. Repeatability over the dynamic range was determined by the coefficient of variation (cv) of the measured target copy number or of the MON810 content between the replicates of the dilution series. 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]. Bread and pastry is part of our everyday life. This collection of research titles about culinary consists of every topic from food poisoning to preservation methods. Thus, a simple ddPCR testing set-up is proposed (set up B, Table S9 in Appendix S2) that included two replicate reactions for each test portion of the sample, in accordance with the ISO 21570:2005 standard [30]. Just fill out a simple form. Moreover, its use for several types of food, feed and seed matrices commonly found in routine samples has been verified. In the same research determine th. To assess trueness, data generated by the experiments for dynamic range determination and for intra- and inter-cartridge repeatability performance were used. One example of the need for quantitative nucleic acid analysis in food and feed is the testing for genetically modified organisms (GMOs). Qualitative Methods in Food Related Research Lauren Blake Royal Veterinary College / LCIRAH ANH Academy Week June 2016. In the absence of a DNA reference material certified for absolute copy number concentration, trueness could only be assessed for the MON810 content. Several MON-ØØ81Ø-6 (MON810) maize seed powder based, certified reference materials (CRM) were purchased from the EU Joint Research Centre, IRMM (Institute for Reference Materials and Measurements, Geel, Belgium). 2 - The American supermarket: Stuck in a timewarp? 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. Fast food vs. slow food: what will be the trend in the future? No, Is the Subject Area "Eating habits" applicable to this article? Salmonella cases in port cities: a quantitative evaluation. Both types can be used to start exciting research. In parallel, DNA from sample G147/08 was also extracted and purified according to a modified CTAB method (see Appendix S1) [21]. Ph.D. or Masters Students can complete research with the help of the given food and nutrition research paper topics suggested by Students Assignment Help. How do restaurant services manage the Big 8 allergens? 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. From these results, it was concluded that the duplex ddPCR assay performs as well as the singleplex ddPCR assays without any additional optimization. Results from the dilution series used for the dynamic range, the aLOD and aLOQ determination, and the overall repeatability. No, Is the Subject Area "Genetically modified organisms" applicable to this article? When writing a research title, you can use the four criteria listed above as a guide. The aim of your quantitative research is to determine the number of students in your High school taking up TVL Home Economics now, are planning to study food and beverage courses at college and will pursue a career in the food and beverage industry. Food and cookery is a wide area, which means it includes all the issues related to food. We thank Prof. Dr. Roger Pain for reviewing the manuscript. These experiments demonstrate that, using ddPCR, one can obtain repeatable and comparable quantitative estimates of GMO target number or content. Funding: The work was supported by the Slovenian Research Agency (grant Nos. That’s why they come to Mintel. Is the atmosphere that important for restaurants? A limited specificity study was however conducted on two samples containing either wild-type maize, or DNA from milk sample without maize. Healthy fast-food chains in the US: an overview. 7 - Gluten-free: Is the growth sustainable? How are robots used in the restaurant industry? Another important factor when introducing new methods or technologies for testing GMOs in food and feed is their applicability. 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. PLoS ONE 8(5): What are the best oils for frying: saturated fat issue? This food survey questions template helps to evaluate food quality and food satisfaction feedback from the customers. https://doi.org/10.1371/journal.pone.0062583.s001, https://doi.org/10.1371/journal.pone.0062583.s002, https://doi.org/10.1371/journal.pone.0062583.s003. If you are more interested in a practical qualitative approach, here is a great list of qualitative research titles about cookery: Food and cookery is a wide area, which means it includes all the issues related to food. Feel free to alter any topic to your liking. This book focuses on the food safety challenges in the vegetable industry from primary production to consumption. Why are cereals becoming the breakfast of the past? An attempt to optimize the duplex qPCR assay was made by varying primers and probe concentration of both systems. 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. All samples used for determining the dynamic range came from serial dilution of a single stock MON810 maize DNA sample. How to use wines for cooking French cuisine properly? If you are not sure how to choose a food research topic, here is a short guide for you. https://doi.org/10.1371/journal.pone.0062583.g003. Both stock and diluted DNA solutions from the CTAB extract could be used for MON810 content determination with qPCR. It has already been asserted that qPCR offers a much broader dynamic range than digital PCR [16]. 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:. In addition, for all experiments performed in this study, NTC resulted in no signal (no positive droplet). The overall repeatability could also be estimated by analyzing the results of the experiment performed for aLOQ and dynamic range determination. The impact of a visual representation of a dish on a customer’s review. The food and restaurant market is a vast and demanding industry that is expanding at a rapid pace. Unfortunately, your browser is too old to work on this site. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. More specifically, their ability to perform well with different sample matrices and within a range of concentrations relevant for GMO testing has to be demonstrated [24]. Real-time quantitative polymerase chain reaction (qPCR) is currently used for quantitative molecular analysis of the presence of GMOs in products. This is because many students do not prepare well for the food research projects, owing to low analytical and quantitative research abilities to search for a relevant topic for research topics on food and cookery. Yes If more samples must be handled simultaneously, ddPCR shows even better throughput and smaller cost than qPCR (see Tables S11 and S12 in Appendix S2). And it gives unlimited career opportunities for those interested in the art of cooking. Yes 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]. When quantitative analysis is required, the use of real-time quantitative PCR (qPCR) is preferred because of its accuracy and precision [1]. 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. Even if the issue has been researched, you can quickly narrow it down. Yes 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.