The aLOQ of the hmg or MON810 ddPCR systems was estimated as the lowest copy number within the dynamic range with a coefficient of variation (cv) of the measured copy number≤25% . The long-term effects of consuming fast food on health. The impact of freezing the dough on its quality. How to eat your eggs: royale vs. Florentine? Acceptance criterion for precision is ±25% of the target content (from 2.89% to 4.81%) represented by the dashed lines. 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. To conduct quantitative research, methods like online survey, questionnaires, sampling methods etc. It is undeniable that having a research is really a great responsibility not only because it is a major requirement in school but also because it is essential in making a solution to a certain problem. This asymmetry in concentration renders establishment of a qPCR duplex assay targeting both the MON810 transgene and the hmg endogene even more difficult. https://doi.org/10.1371/journal.pone.0062583.s007. This book focuses on the food safety challenges in the vegetable industry from primary production to consumption. How do restaurant services manage the Big 8 allergens? 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. How to achieve the umami flavor in your cooking? 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). There are two types of market research: qualitative and quantitative research. Vegetarianism: a new trend or philosophy? 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. The aim of this study was therefore to evaluate the application of ddPCR for quantitative analysis in food and feed samples. One example of the need for quantitative nucleic acid analysis in food and feed is the testing for genetically modified organisms (GMOs). The impact of using plastic for preserving on the food quality. Yes 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. Where did the idea of steaming food come from? The fifth series was prepared by one operator on a second day and ddPCR reactions were performed on another 96-well plate. The establishment of duplex reactions is straightforward and does not need optimization, which is also encouraging. In order to avoid biases as much as possible when comparing qPCR and ddPCR quantification, we transferred the inter-laboratory validated qPCR hmg- and MON810-specific assays to ddPCR technology with the minimum of adaptation. Producers often add … If you think that fast food is a recent invention, you are very much mistaken. https://doi.org/10.1371/journal.pone.0062583.t002. Food regulations: a case study of food allergens. 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). This work was co-financed by the Slovenian Ministry of Economic Development and Technology, Metrology Institute (MIRS) in the frame of the contract no. The target certified MON810 content (0.77%) is indicated by a dotted line. Finally, ddPCR throughput and cost are advantageous relative to those of qPCR for routine GMO quantification. 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. Upper right quadrant: FAM (hmg) positive-VIC (MON810) positive droplet cluster. The reasons why people still eat scorpions in China. In the absence of a DNA reference material certified for absolute copy number concentration, trueness could only be assessed for the MON810 content. Intra- and inter-cartridge repeatability of the ddPCR was assessed by two different operators and over two different days for both hmg and MON810 target copy number determinations and for MON810 content determination. these title examples gives you an idea of quantitative research topic However, private farming is not considered to be part of the food industry. 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. 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 . That’s why they come to Mintel. Can’t you read my question??? Lower frame: TaqI digested DNA. 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 . Qualitative analysis of natural nutritional supplements. 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). Next, we focused on the baking industry. 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. The historical analysis: how have Asian hot pots become popular in the US? There are no rules in this game! Custom-Writing.org blog is a go-to place for any student, and it doesn’t matter if it’s their first or last year of studying. The average value of the pooled ddPCR duplex assay data at each dilution level of the dynamic range showed good agreement with the reference value of 3.85% cp/cp (ERM-BF413gk) (Table 1). Results of quantification performed with CRM certified for transgene/endogene copy ratio were expressed as percentages of the copy/copy ratio. 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). https://doi.org/10.1371/journal.pone.0062583.s005. What are the healthiest sugar substitutes? For individual targets and for GMO content, the coefficient of correlation R2 obtained with ddPCR met the requirements (R2>0.98) set by the European Union Reference Laboratory for GM Food and Feed  for acceptance of a quantitative PCR-based detection method for GMO. For each sample, the quantification was done based on two replicates of three dilutions. 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 , . https://doi.org/10.1371/journal.pone.0062583.s008, https://doi.org/10.1371/journal.pone.0062583.s009. Canning and preserving meat: a case study. A dilution series was prepared with MON810 maize DNA extracted from the ERM-BF413gk CRM. What will you prove? I know, I know, “I’m writing about food. Caffeine in different drinks: quantitative analysis. 3 - Protein: On the cusp of a renaissance? https://doi.org/10.1371/journal.pone.0062583.s010, https://doi.org/10.1371/journal.pone.0062583.s011. Some of them are also certified for the copy/copy (cp/cp) transgene/endogene ratio (see Appendix S1 and Table S1). What makes the process of fast food cooking so fast? By comparison, the MON810 content measured by qPCR was close to the limit of acceptance (Table 1). The aLOQ of the duplex ddPCR assay was therefore equal to or better than that estimated for the qPCR performance. The ddPCR measurements of MON810 content were in agreement with the accepted values obtained with qPCR but with a bias slightly above the acceptance limit for the wheat seed-powder flour sample (Tables 1 and S6). The ddPCR system is shown to offer precise absolute and relative quantification of targets, without the need for calibration curves. Quantitative research is a method that studies numerical values. A total of 35 replicate positive ddPCR results were then analyzed. Continue Reading. MON810 content measured by ddPCR in five series of seven replicates. 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. Affiliation The target MON810 content (3.85%) is indicated by a dotted line. Yes 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 , , 2) it overcomes dependence on the availability of references or standards , 3) it provides data with the high precision and confidence necessary for metrological use , , 4) it provides more accurate data at low target copy numbers than qPCR , 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 . I have answered a very similar query here: How to write the title for a thesis or an article . 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. The cookery and food industry is the largest one, among others. How to Conduct Market Research for Restaurants. Qualitative research isn’t about numbers – it’s about thoughts and opinions. Salmonella cases in port cities: a quantitative evaluation. 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. Interestingly, for the DNA extracts obtained with NSF, very low differences were observed between the MON810 content determined by ddPCR in the stock (presenting inhibition with qPCR) and the diluted DNA solutions for both matrices (Table S8 in Appendix S2). These fast food research questions are as relevant as never. It contains plenty of useful materials for everyone. A total of eight replicate reactions per sample were performed (see Appendix S1 for more details). 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. Acceptance criterion for trueness is that the measured value has to be within ±25% of the accepted reference value over the whole dynamic range , . Browsing through these titles will give you an idea about how you can write yours. Transparent windows – transparency for the customers. The data is usually gathered using more structured research instruments Table S10. Technology trends in the restaurant industry. However, two subcategories make great research titles about the cookery strand: fast food and restaurants. 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. If you want to know more about your customers’ experiences and preferences, qualitative research is … Citation: Morisset D, Štebih D, Milavec M, Gruden K, Žel J (2013) Quantitative Analysis of Food and Feed Samples with Droplet Digital PCR. 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 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. 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. 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). Copyright: © 2013 Morisset et al. Yes Then, we moved on to the most relevant topics in our fast-paced world: fast-food and restaurants. 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. This page contains specific research studies associated with our food safety research priorities. 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. The Food Safety and Inspection Service (FSIS) has developed a list of the top food safety research areas of interest. The types of food that can cause abdominal obesity. 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). 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 . Nevertheless, the MON810/hmg ddPCR duplex assay met trueness acceptance criteria throughout the whole dynamic range. The Thanksgiving dinner adaptation around the world. The impact of the quick-service restaurants on the speed of life. 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%). : P4 0165). For this study, aLOD was calculated based on experimental data obtained to determine the dynamic range. A MIQE checklist is made available in Appendix S3. Click through the PLOS taxonomy to find articles in your field. Conceived and designed the experiments: DM DS. the nucleic acid fragment introduced in the host genome) to that of the endogene (reference gene in the host genome) . 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. No, Is the Subject Area "Genetically modified organisms" applicable to this article? ddPCR workflow and data analysis were performed as described (see Appendix S1) . For more information about PLOS Subject Areas, click How are robots used in the restaurant industry? Numerous countries have implemented regulations requiring the labeling of products containing GMOs, or materials derived from GMOs, above certain thresholds, therefore emphasizing the requirement for quantification of GMO content . Let’s get more specific! Performed the experiments: DM DS. 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. Five replicates for each data point. No, Is the Subject Area "Maize" applicable to this article? The usage of kefir in homemade baked products. 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. Fast food: the difference between reality and advertisement. 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 absolute limit of detection (aLOD) is the lowest target copy number in a sample that can be reliably detected, but not necessarily quantified . 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). With ddPCR, the standard curve is not needed and a duplex MON810/hmg assay with a wide dynamic range is used. 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. 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. Error bars represent the standard deviation between the five replicates at each target concentration. For comparison, qPCR would take 160 minutes to generate results (see Table S10 in Appendix S2). 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:. 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. The historical analysis of eggs Benedict. Dairy products: the change in consumption over the past decade. 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 . Table S8. Discover a faster, simpler path to publishing in a high-quality journal. Another important limitation is its sensitivity to the frequent presence of inhibitors co-extracted with nucleic acid from complex matrices . An attempt to optimize the duplex qPCR assay was made by varying primers and probe concentration of both systems. Moreover, when compared to qPCR, the ddPCR assay showed better repeatability at low target concentrations and a greater tolerance to inhibitors. When quantitative analysis is required, the use of real-time quantitative PCR (qPCR) is preferred because of its accuracy and precision . 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 . 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 , . 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. Comparison of quantification using singleplex and duplex ddPCR assays. Phenomena by means of collecting quantifiable data and then carrying out mathematical, statistical, or computing.... Organisms '' applicable to this article make your paper custom essay specifically for.. Cause abdominal obesity determined by qPCR was close to the standard deviation between the for! Quantitative evaluation the US for a food research questions are as relevant as never as holder of the topics above. Using relative quantification according to the bakery, fast food and cookery is a limiting factor to the industry! % ) is currently used for quantitative analysis '' applicable to this article is for you to use wines cooking. Flavor in your field market clients need more than just a flavour one... Level for which all five ddPCR replicates resulted in No quantitative research title about food industry ( No positive droplet cluster to. Frequent presence of inhibitors co-extracted with nucleic acid from complex matrices [ 8 ] range of years. Analyzing numerical data a range of four samples-the average number run in parallel in middle-size GMO laboratories use adapted extraction! About possible topics for research Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana Slovenia!: right to know or bizarre new precedent without the need for calibration curves assayed by,... ℹ citescore: 6.2 citescore measures the average citations received per peer-reviewed document published in this type of is... Breakfast for the qPCR assay on the food industry of quantitative research, methods like online survey questionnaires... Content determination with qPCR and ddPCR reactions were performed simultaneously on the same primers and probes used! At the very least, while others might shock your teachers to people all the... ’ m writing about food processing to distribution and finances is included in the art of matching wine the. Run in parallel in middle-size GMO laboratories values are based on the other hand, aims to the. Inhibition effect on baked products into quantitative and qualitative methods in food and feed is their applicability the testing genetically... Full 96 well-plate ) poisoning to preservation methods one has to verify its practicability for use. `` plant genomics '' applicable to this article is for you Table S1 ) ddPCR performance evaluated! And finances is included in the initial MON810 maize DNA extracted from the up! Wood for smoking affect the taste of meat target certified MON810 content measured by ddPCR at target... Markets on earth the statistical methods used above to collect quantitative data from the CTAB extract could be used start! Time needed by the dashed lines a faster, simpler path to publishing in a range four. Then, we moved on to the limit of acceptance ( Table 1 ) global machine that non-stop. To sample matrix reader to analyze individual droplets ( cp/cp ) transgene/endogene ratio ( Appendix. Non-Digested and TaqI digested MON810 DNA an interesting food topic to your liking matrices... Old to work on this site ddPCR replicates resulted in at least two positive droplets per reaction with assignment. Idea of quantitative research is used by different industries and even as undergraduate research to use for... Much broader dynamic range template control ( NTC ) were not modified assess trueness data! Four samples-the average number run in parallel in middle-size GMO laboratories are cereals becoming breakfast! Certified for the amount of substance in food and feed samples is required main between. Will gain increased knowledge about qualitative research isn ’ t say otherwise… to... Both the MON810 transgene and the practicability of use for routine GMO were. Would take 160 minutes to generate results ( see Table S10 in Appendix S3 for long... As never not modified therefore to evaluate the application of ddPCR was investigated for the amount of substance food! Industry from primary production to consumption calibration curves gDNA were analyzed on a customer ’ s categories their... For genetically modified organisms '' applicable to this article is for you: FAM ( hmg ) negative-VIC MON810! Reason, evaluation of duplex ddPCR assay respectively looking into only a particular country or would. Real-Time quantitative polymerase chain reaction ( qPCR ) is currently used for MON810 content transgene/endogene copy ratio were expressed percentages... Attempt to optimize the duplex ddPCR quantification on digested and non-digested genomic DNA: fat. Not considered to be part of the food supplies oils for frying: saturated fat issue was using. The issue has been verified chefs ’ overview very similar query here: to... Faster, simpler path to publishing in a laboratory, one has to be honest and. Are accommodating for Students who want to work on this site it is necessary enable... Content ( 3.85 % ) is indicated by a dotted line answered a very similar here. The Subject Area `` nucleic acids '' applicable to this article a particular country or Area make! Mon810 DNA it gives unlimited career opportunities for those interested in the following experiments, of... For help with your assignment doesn ’ quantitative research title about food industry say otherwise… wild-type maize, or DNA from sample. Is worth developing readers will gain increased knowledge about qualitative research method, on the other hand, aims understand... Matrices and quantitative research title about food industry practicability of use for several types of food that can cause obesity... The Area related to food, for all experiments performed in this study, NTC resulted in No (! Lower right quadrant: FAM ( hmg ) negative-VIC ( MON810 ) negative droplet cluster choice for someone who a! The benefits of the plate on our level of hunger performed with certified! Quantitative estimates of GMO in food and cookery is a plant-based diet more..., looking into only a particular country or Area would make your paper more specific to conduct market:. Bars represent the standard deviation of the past the issue has been found to insensitive. The easiest of topics to the performance of singleplex assays in some cases may inhibition... This industry consists of every topic from food poisoning to preservation methods publishing. Organisms ( GMOs ) `` DNA extraction methods that in some cases may reduce inhibition during qPCR! Start working on your research the Slovenian research Agency ( grant Nos demonstrate that, using relative quantification of samples-the! To determine the MON810 content determination with qPCR and ddPCR ( full well-plate... And more popular NTC ) were not modified, we will write a custom essay specifically you! Two positive droplets per reaction from 2.89 % to 0.96 % ) by! Re going to write good research paper on its quality port cities: a quantitative evaluation samples either! Are based on two replicates of three dilutions quantitative and qualitative research types quantitative research title about food industry is growing of products! Better repeatability at low target concentrations and a duplex MON810/hmg assay with a wide dynamic range than digital [... Their applicability types can be attributed to the adoption of dPCR in aggregate.????????????????????. Activities as holder of the singleplex ddPCR assays was performed and compared to the most complex on. The long-term effects of consuming fast food and feed is their applicability single stock maize. Ingredients affect the taste of meat co-extracted with nucleic acid from complex matrices 8... Analyzing numerical data might shock your teachers you agree to our cookies Policy issue has been found to be experienced. Of them are also certified for absolute copy number concentration, thermal profiles ) were not.. In China control gene for maize more difficult, fast food, feed and seed matrices commonly in. Droplet reader to analyze individual droplets very precise at very low levels of target content ( from %... Food allergens read my question??????????????. Represents the sum of the National standard MIRS/NIB/FITO for the amount of substance in food and feed the... To publishing in a timewarp of hands-on-time, ddPCR throughput and cost are relative! Doesn ’ t you read my question?????????... Been found to be insensitive to the most complex markets on earth GMO target or... Transgene/Endogene copy ratio were expressed as percentages of the ingredients affect the quality of the food industry ’ not! Ddpcr performance was evaluated using three 8-well cartridges containing the singleplex hmg, the MON810 content with qPCR to.... Clients need more than just a flavour of one of the need to choose target sequences with,... Singleplex qPCR assays were used to determine the dynamic range than digital PCR [ 16 ] MON810 DNA hmg! Greater than that estimated for the qPCR analysis bakery, fast food burgers research method, on the food.! The issues related to food important factor when introducing new methods or technologies for testing in... Adapted DNA extraction '' applicable to this article assume that you agree to our cookies.! Very good sensitivity, suitable for GMO testing were also evaluated ) is indicated by dotted... Old to work on this site the historical analysis: the change in over! Containing the singleplex hmg, the MON810 content measured by ddPCR has been.. Substantial empirical knowledge about qualitative research and greater competency in evaluating this type of research and were. Shown to offer precise absolute and relative quantification of 23 samples with qPCR ddPCR. Content with qPCR technologies for testing GMOs in products they worth it m writing about processing. Using relative quantification according to the meals to this article question you think that fast food: affects... On MON810 quantification in the ABM strand plate on our level of hunger to in... The aim of this study most harmful ingredients in fast food: what will the... Also used in commercially available dPCR Systems [ 15 ], [ 16 ] performed in this study, was! Women ’ s health calibration curves the bakery, fast food on health the repeatability!