The absolute sensitivity is, according to our own assessment (data not shown), lower than the one observed for the MON810 singleplex qPCR assay at around 6–18 copies, and comparable to the one observed in cdPCR . Evaluation of qPCR duplex systems has shown that the hmg system performed identically in duplex and singleplex qPCR reactions, while MON810 amplification was significantly affected in the duplex reactions, showing signal values approximately 5.5 Cq higher than in singleplex reactions (data not shown). Lower left quadrant: FAM (hmg) negative-VIC (MON810) negative droplet cluster. The amount of fats children and adults need: statistical analysis. https://doi.org/10.1371/journal.pone.0062583.g002. Our food market clients need more than just a flavour of one of the most complex markets on earth. A qualitative research method, on the other hand, aims to understand why people think and behave a certain way. 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. Results of quantification performed with CRM certified for transgene/endogene copy ratio were expressed as percentages of the copy/copy ratio. In the same research determine th. Next, we focused on the baking industry. Both stock and diluted DNA solutions from the CTAB extract could be used for MON810 content determination with qPCR. Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia. For qPCR, measurements were made in duplicate. Let’s get more specific! MON810 content measured by ddPCR in five series of seven target concentrations. I have answered a very similar query here: How to write the title for a thesis or an article . For example, looking into only a particular country or area would make your paper more specific. 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 , . How do different types of flour affect the quality of the cake? https://doi.org/10.1371/journal.pone.0062583.s008, https://doi.org/10.1371/journal.pone.0062583.s009. Caffeine in different drinks: quantitative analysis. Eating disorders and beauty standards: a quantitative evaluation. Based on this criterion, aLOQ was estimated to be around 5 copies for the hmg system, and 18 copies for the MON810 system and for the duplex ddPCR assay (Table S5). This asymmetry in concentration renders establishment of a qPCR duplex assay targeting both the MON810 transgene and the hmg endogene even more difficult. It is also very precise at very low levels of target content. However, private farming is not considered to be part of the food industry. aLOD was determined as the lowest concentration level for which all five ddPCR replicates resulted in at least two positive droplets per reaction. Fast food: the difference between reality and advertisement. Set-up and number of reactions needed for quantification with qPCR and ddPCR (full 96 well-plate). 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. 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. What are the most harmful ingredients in fast food burgers? The data is usually gathered using more structured research instruments 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. The cookery and food industry is the largest one, among others. Coffee for breakfast: a destructive habit for women’s health. With ddPCR, the standard curve is not needed and a duplex MON810/hmg assay with a wide dynamic range is used. 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. What makes the process of fast food cooking so fast? They are accommodating for students who want to write a paper related to food. In this research method, researchers and statisticians deploy mathematical frameworks and theories that pertain to … It was recently discussed as to whether price is a limiting factor to the adoption of dPCR in the laboratory . An additional set of experiments was performed to establish repeatability between different emulsification runs. That’s why they come to Mintel. Since inhibition is assessed during the screening and identification phases of GMO testing and given the demonstrated tolerance of the assay for amplification inhibition, it is not necessary to control the inhibition at the ddPCR quantification stage. 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 . Consequently, diluted DNA samples were used to determine the MON810 content with qPCR. The long-term effects of consuming fast food on health. In this study, similar variability of the measured target copy numbers (Table S5 and Figure 1) and of the MON810 content was observed at lower target concentrations (Table S5 and Figure 3). All along the dynamic range, the cv of the determined hmg copies, MON 810 copies, and MON810 content remained below the threshold for acceptance of quantitative methods (cv<25%) (Table S5). After end-point PCR amplification, each partition is scrutinized and defined as positive (“1”, presence of PCR product) or negative (“0”, absence of PCR product) hence the term “digital”. The historical analysis: the real birthplace of curry. This collection of research titles about culinary consists of every topic from food poisoning to preservation methods. Linearity of response, absolute limits of detection and quantification, repeatability over the dynamic range of the ddPCR endogene and transgene assays were assessed. Chemicals of Potential Concern In the absence of a DNA reference material certified for absolute copy number concentration, trueness could only be assessed for the MON810 content. An attempt to optimize the duplex qPCR assay was made by varying primers and probe concentration of both systems. Wrote the paper: DM MM KG JZ. https://doi.org/10.1371/journal.pone.0062583, Editor: Joshua L. Heazlewood, Lawrence Berkeley National Laboratory, United States of America, Received: January 29, 2013; Accepted: March 22, 2013; Published: May 2, 2013. It has already been asserted that qPCR offers a much broader dynamic range than digital PCR . 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. RESEARCH PROPOSAL Title: A quantitative study of the attitude, knowledge and experience of staff nurses on prioritizing comfort measures in care of the dying patient in an acute hospital setting. Yes During qPCR tests, we detected the presence of inhibition in the stock DNA solution of two samples (wheat seed-powder flour with maize contamination and maize feed), as indicated by the large differences in MON810 content (cv>25%) calculated from different sample dilutions. 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 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). By comparison, the MON810 content measured by qPCR was close to the limit of acceptance (Table 1). Discover a faster, simpler path to publishing in a high-quality journal. Is the atmosphere that important for restaurants? You can undoubtedly find a question you think is worth developing. Five replicates of the dilution series and of a non template control (NTC) were measured by ddPCR. This industry consists of every process connected to producing and selling food. Further, ddPCR performance was evaluated using non-digested gDNA. No, Is the Subject Area "Quantitative analysis" applicable to this article? Quantification of routine samples using ddPCR is therefore practical and has the potential to provide better throughput and cost-effectiveness than qPCR for GMO laboratories. 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. Other samples containing the MON810 maize event, and previously assayed by qPCR, were also used in this study. 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. The lack of certified reference material has also been noted . Why do people prefer organic milk: qualitative research? 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). 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). these title examples gives you an idea of quantitative research topic CiteScore: 6.2 ℹ CiteScore: 2019: 6.2 CiteScore measures the average citations received per peer-reviewed document published in this title. https://doi.org/10.1371/journal.pone.0062583.s010, https://doi.org/10.1371/journal.pone.0062583.s011. Ask our academic expert for help with your assignment! In our laboratory, the aLOQ estimated for the qPCR assay on the same DNA is 18 copies. Epistemologists refer to this as quantitative research (1, 2, 3).It is well suited to testing the effectiveness of interventions, evaluating outcomes, mining statistical … Restaurant industry: smaller plates trend. https://doi.org/10.1371/journal.pone.0062583.s007. There is a list of food-related topics for a research paper below. Caffeine dependency in Italy: ethnographic research. People hate waiting for their food for too long, so the fast-food demand is growing. A limited specificity study was however conducted on two samples containing either wild-type maize, or DNA from milk sample without maize. Fast food vs. slow food: what will be the trend in the future? These experiments demonstrate that, using ddPCR, one can obtain repeatable and comparable quantitative estimates of GMO target number or content. Moreover, the key assay parameters (primer and probe concentration, thermal profiles) were not modified. For ddPCR, no significant variation of the measured target copy number was observed between the singleplex and duplex ddPCR assays for both hmg (bias = −1.8%) and MON810 systems (bias = 3.7%). 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). No, Is the Subject Area "Eating habits" applicable to this article? This book focuses on the food safety challenges in the vegetable industry from primary production to consumption. 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). How to eat your eggs: royale vs. Florentine? Trueness is defined as the closeness of agreement between the average value obtained from a series of test results and an accepted reference value . Salmonella cases in port cities: a quantitative evaluation. 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 average number of droplets read for each ddPCR reaction included in the data analysis was 13,606 with a standard deviation of 931 droplets (coefficient of variation cv = 6.8%). In summary, the ddPCR MON810/hmg duplex assay can be applied for routine quantification of the MON810 maize, as demonstrated on a large range of transgene content (experimentally from 0.04% to 29.6%) usually found in samples. MON810 content was determined by qPCR, using relative quantification according to the standard curve approach. It is so easy to make a first writing step. VIC vs. FAM channel clustering plot of droplets for non-digested and TaqI digested MON810 DNA. The DNA extracts from samples G053/12 (approximately 6,200 copies of hmg) and G031/12 (approximately 100 ng DNA) were tested with the duplex MON810/hmg ddPCR assay. 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. It also shows very good sensitivity, suitable for GMO testing. The absolute limit of quantification (aLOQ) and absolute limit of detection (aLOD) for qPCR and ddPCR were determined on these experimental results. However, if you don’t want to waste your time looking for the best topic, this article is for you! The main difference between the two approaches can be attributed to the time needed by the droplet reader to analyze individual droplets. Is the Subject Area "Polymerase chain reaction" applicable to this article? Set-up and number of reactions needed for simultaneous quantification of 23 samples with qPCR and ddPCR. Browsing through these titles will give you an idea about how you can write yours. 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 , , ) and of point mutations. How are robots used in the restaurant industry? 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