{"title":"Molecular Markers for the Evaluation of Clonal Fidelity in Medicinal Plants","authors":"","doi":"10.2174/9789811439315120010018","DOIUrl":"https://doi.org/10.2174/9789811439315120010018","url":null,"abstract":"Medicinal plants are major sources of secondary metabolites for which they\u0000have been paid more attention by pharmaceutical industries. In order to produce these\u0000secondary metabolites, medicinal plants are cultivated and for that plant tissue or\u0000organ, culture can be a suitable alternative. However, these plants are treated with plant\u0000hormones and elicitors to enhance the secondary metabolites and such elicitation may\u0000lead to genetic or epigenetic changes which are known as somaclonal variations. Thus,\u0000a stringent method of monitoring is required to observe the true-to-types of these\u0000medicinal plants when multiplied through tissue culture. Molecular markers like\u0000Randomly Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeat\u0000(ISSR), and Simple Sequence Repeats (SSR) are highly suitable markers to assess\u0000clonal fidelity in micropropagated medicinal plants. In the present chapter, the\u0000execution of such markers to check somaclonal variations in tissue culture raised\u0000medicinal plants is discussed in detail.","PeriodicalId":179247,"journal":{"name":"Protocols used in Molecular Biology","volume":"26 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123193534","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"List of Contributors","authors":"N. Beopoulos","doi":"10.2174/9789811439315120010003","DOIUrl":"https://doi.org/10.2174/9789811439315120010003","url":null,"abstract":"","PeriodicalId":179247,"journal":{"name":"Protocols used in Molecular Biology","volume":"25 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125159190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The Recent Advancement in Rapid Golgi Method and Result Interpretation","authors":"","doi":"10.2174/9789811439315120010017","DOIUrl":"https://doi.org/10.2174/9789811439315120010017","url":null,"abstract":"The foundation knowledge of recent advancements of neuroscience was\u0000based on the Golgi staining observations. This is one of the best approaches to visualise\u0000the neuronal cytoarchitecture and complete morphology of neurons with incomparable\u0000clarity. This technique is based on the principle of heavy metal impregnation. There are\u0000many modifications and advancement occurred to improve the visualization. This\u0000chapter will provide the recently used protocols to visuals the neuronal architecture,\u0000dendritic arborization and spine density in different brain regions. Along with the\u0000manual observation, the present chapter also describes the currently used tools and\u0000software for the better understanding and visualisation of neurons.","PeriodicalId":179247,"journal":{"name":"Protocols used in Molecular Biology","volume":"473 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116516315","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Analyzing Gene Expression through Real Time PCR while Neo-tissue Regeneration using Developed Tissue Constructs","authors":"","doi":"10.2174/9789811439315120010006","DOIUrl":"https://doi.org/10.2174/9789811439315120010006","url":null,"abstract":"Real-time PCR offers a wide area of application to analyze the role of gene\u0000activity in various biological aspects at the molecular level with higher specificity,\u0000sensitivity and the potential to troubleshoot with post-PCR processing and difficulties.\u0000With the recent advancement in the development of functional tissue graft for the\u0000regeneration of damaged/diseased tissue, it is effective to analyze the cell behaviour\u0000and differentiation over tissue construct toward specific lineage through analyzing the\u0000expression of an array of specific genes. With the ability to collect data in the\u0000exponential phase, the application of Real-Time PCR has been expanded into various\u0000fields such as tissue engineering ranging from absolute quantification of gene\u0000expression to determine neo-tissue regeneration and its maturation. In addition to its\u0000usage as a research tool, numerous advancements in molecular diagnostics have been\u0000achieved, including microbial quantification, determination of gene dose and cancer\u0000research. Also, in order to consistently quantify mRNA levels, Northern blotting and in\u0000situ hybridization (ISH) methods are less preferred due to low sensitivity, poor\u0000precision in detecting gene expression at a low level. An amplification step is thus\u0000frequently required to quantify mRNA amounts from engineered tissues of limited size.\u0000When analyzing tissue-engineered constructs or studying biomaterials–cells\u0000interactions, it is pertinent to quantify the performance of such constructs in terms of\u0000extracellular matrix formation while in vitro and in vivo examination, provide clues\u0000regarding the performance of various tissue constructs at the molecular level. In this chapter, our focus is on Basics of qPCR, an overview of technical aspects of Real-time\u0000PCR; recent Protocol used in the lab, primer designing, detection methods and\u0000troubleshooting of the experimental problems.","PeriodicalId":179247,"journal":{"name":"Protocols used in Molecular Biology","volume":"6 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130247963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Culture and Maintenance of Human Ovarian Carcinoma Cells for Scrutinizing Anti-cancerous Activities of Various Compounds via Some Potent Molecular Markers","authors":"","doi":"10.2174/9789811439315120010015","DOIUrl":"https://doi.org/10.2174/9789811439315120010015","url":null,"abstract":"Ovarian carcinoma is the 5th most common type of cancer of gynecologic\u0000origin and accounts for about one-fourth of the total malignancies of the female genital\u0000tract. Ovarian carcinoma accounts for highest mortality in females due to the\u0000development of chemo-resistance against drugs and lack of symptoms and undetectable\u0000biomarkers in the early stages of diagnosis. Tumour debulking, chemotherapies,\u0000radiotherapies, targeted therapies, immunotherapies and stem cell transplants are some\u0000of the measures that have been adopted by the experts for curing the disease but still,\u0000full control over the problem has not been achieved. Research on various herbal and\u0000chemosynthetic nano-compounds have shown a new light in this regard, as the studies\u0000on them so far have revealed that they have anti-proliferative and apoptotic properties\u0000that will help in finding new ways to develop drugs for cancer patients. This chapter\u0000deals how to culture and maintain the human ovarian carcinoma cell lines in the\u0000laboratory which are being procured from cell repositories and then to study the anticancer\u0000efficacy of various promising compounds by potent molecular markers like cellcycle\u0000progression and annexin V- FITC apoptosis detection.","PeriodicalId":179247,"journal":{"name":"Protocols used in Molecular Biology","volume":"3 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126116430","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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