{"id":29345,"date":"2022-12-29T09:20:19","date_gmt":"2022-12-29T09:20:19","guid":{"rendered":"https:\/\/evaggelatos.com\/?p=29345"},"modified":"2022-12-29T09:32:59","modified_gmt":"2022-12-29T09:32:59","slug":"the-use-of-pcr-as-an-educational-guide-for-cellular-engineering","status":"publish","type":"post","link":"https:\/\/evaggelatos.com\/?p=29345","title":{"rendered":"The use of PCR as an educational guide for cellular engineering (\u03ba\u03b1\u03b9 \u03bc\u03b5\u03c4\u03ac\u03c6\u03c1\u03b1\u03c3\u03b7 \u03c3\u03c4\u03b1 \u03b5\u03bb\u03bb\u03b7\u03bd\u03b9\u03ba\u03ac)."},"content":{"rendered":"<h1 class=\"heading-title\">Molecular cloning using polymerase chain reaction, an educational guide for cellular engineering<\/h1>\n<div class=\"inline-authors\">\n<div class=\"authors\">\n<div class=\"authors-list\"><span class=\"authors-list-item \"><a class=\"full-name\" href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Hoseini+SS&amp;cauthor_id=25745516\" data-ga-category=\"search\" data-ga-action=\"author_link\" data-ga-label=\"Sayed Shahabuddin Hoseini\">Sayed Shahabuddin Hoseini<\/a><sup class=\"affiliation-links\"><span class=\"author-sup-separator\">\u00a0<\/span><a class=\"affiliation-link\" title=\"Departments of Pediatric Hematology, Oncology and Blood Stem Cell Transplantation, Hannover, Germany ; Hannover Center for Transplantation Research, Hannover, Germany.\" href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/25745516\/#affiliation-1\"> 1 <\/a><\/sup><span class=\"comma\">,\u00a0<\/span><\/span><span class=\"authors-list-item \"><a class=\"full-name\" href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Sauer+MG&amp;cauthor_id=25745516\" data-ga-category=\"search\" data-ga-action=\"author_link\" data-ga-label=\"Martin G Sauer\">Martin G Sauer<\/a><sup class=\"affiliation-links\"><span class=\"author-sup-separator\">\u00a0<\/span><a class=\"affiliation-link\" title=\"Departments of Pediatric Hematology, Oncology and Blood Stem Cell Transplantation, Hannover, Germany ; Hannover Center for Transplantation Research, Hannover, Germany ; Department of Pediatric Hematology and Oncology, Medizinische Hochschule Hannover, OE 6780, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.\" href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/25745516\/#affiliation-2\"> 2 <\/a><\/sup><\/span><\/div>\n<\/div>\n<\/div>\n<div class=\"short-article-details\">Affiliations<\/div>\n<header id=\"heading\" class=\"heading\">\n<div id=\"full-view-heading\" class=\"full-view\">\n<ul id=\"full-view-identifiers\" class=\"identifiers\">\n<li><span class=\"identifier pubmed\"> <span class=\"id-label\"> PMID: <\/span> <strong class=\"current-id\" title=\"PubMed ID\">25745516<\/strong> <\/span><\/li>\n<li><span class=\"identifier pmc\"> <span class=\"id-label\"> PMCID: <\/span> <a class=\"id-link\" href=\"http:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/pmc4350901\/\" target=\"_blank\" rel=\"noopener\" data-ga-category=\"full_text\" data-ga-action=\"PMCID\"> PMC4350901 <\/a> <\/span><\/li>\n<li><span class=\"identifier doi\"> <span class=\"id-label\"> DOI: <\/span> <a class=\"id-link\" href=\"https:\/\/doi.org\/10.1186\/1754-1611-9-2\" target=\"_blank\" rel=\"noopener\" data-ga-category=\"full_text\" data-ga-action=\"DOI\"> 10.1186\/1754-1611-9-2 <\/a> <\/span><\/li>\n<\/ul>\n<p><span class=\"free-label\">Free PMC article<\/span><\/p>\n<\/div>\n<\/header>\n<div id=\"abstract\" class=\"abstract\">\n<h2 class=\"title\">Abstract<\/h2>\n<div id=\"eng-abstract\" class=\"abstract-content selected\">\n<p><strong class=\"sub-title\"> Background: <\/strong> Over the last decades, molecular cloning has transformed biological sciences. Having profoundly impacted various areas such as basic science, clinical, pharmaceutical, and environmental fields, the use of recombinant DNA has successfully started to enter the field of cellular engineering. Here, the polymerase chain reaction (PCR) represents one of the most essential tools. Due to the emergence of novel and efficient PCR reagents, cloning kits, and software, there is a need for a concise and comprehensive protocol that explains all steps of PCR cloning starting from the primer design, performing PCR, sequencing PCR products, analysis of the sequencing data, and finally the assessment of gene expression. It is the aim of this methodology paper to provide a comprehensive protocol with a viable example for applying PCR in gene cloning.<\/p>\n<p><strong class=\"sub-title\"> Results: <\/strong> Exemplarily the sequence of the tdTomato fluorescent gene was amplified with PCR primers wherein proper restriction enzyme sites were embedded. Practical criteria for the selection of restriction enzymes and the design of PCR primers are explained. Efficient cloning of PCR products into a plasmid for sequencing and free web-based software for the consecutive analysis of sequencing data is introduced. Finally, confirmation of successful cloning is explained using a fluorescent gene of interest and murine target cells.<\/p>\n<p><strong class=\"sub-title\"> Conclusions: <\/strong> Using a practical example, comprehensive PCR-based protocol with important tips was introduced. This methodology paper can serve as a roadmap for researchers who want to quickly exploit the power of PCR-cloning but have their main focus on functional in vitro and in vivo aspects of cellular engineering.<\/p>\n<\/div>\n<p><strong class=\"sub-title\"> Keywords: <\/strong> Biological engineering; Cloning; Educational guide; Polymerase chain reaction (PCR); Recombinant DNA; Transduction; Transfection.<\/p>\n<\/div>\n<div id=\"figures\" class=\"figures\">\n<h3 class=\"title\">Figures<\/h3>\n<div id=\"slides-container\" class=\"figures-list\">\n<figure class=\"figure-item \" data-slide-index=\"0\" data-label-slug=\"figure-1\"><a class=\"figure-link\" role=\"button\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig1_HTML.jpg\" aria-describedby=\"figure-caption-0\" data-image-width=\"600\" data-image-height=\"219\" data-image-alt=\"Figure 1\" data-pmc-id=\"PMC4350901\" data-figure-id=\"Fig1\"> <img decoding=\"async\" id=\"article-image-0\" class=\"figure-thumb\" src=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig1_HTML.gif\" alt=\"Figure 1\" \/> <\/a><\/p>\n<div class=\"figure-caption figure-caption-text\" aria-hidden=\"true\">\n<div class=\"caption-wrap\">\n<p>Figure 1<\/p>\n<div class=\"figure-caption-contents\"><b> Overview of the start and\u2026 <\/b><\/div>\n<\/div>\n<\/div>\n<\/figure>\n<figure class=\"figure-item \" data-slide-index=\"1\" data-label-slug=\"figure-2\"><a class=\"figure-link\" role=\"button\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig2_HTML.jpg\" aria-describedby=\"figure-caption-1\" data-image-width=\"600\" data-image-height=\"423\" data-image-alt=\"Figure 2\" data-pmc-id=\"PMC4350901\" data-figure-id=\"Fig2\"> <img decoding=\"async\" id=\"article-image-1\" class=\"figure-thumb\" src=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig2_HTML.gif\" alt=\"Figure 2\" \/> <\/a><\/p>\n<div class=\"figure-caption figure-caption-text\" aria-hidden=\"true\">\n<div class=\"caption-wrap\">\n<p>Figure 2<\/p>\n<div class=\"figure-caption-contents\"><b> Choosing proper restriction enzymes based\u2026 <\/b><\/div>\n<\/div>\n<\/div>\n<\/figure>\n<figure class=\"figure-item \" data-slide-index=\"2\" data-label-slug=\"figure-3\"><a class=\"figure-link\" role=\"button\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig3_HTML.jpg\" aria-describedby=\"figure-caption-2\" data-image-width=\"600\" data-image-height=\"200\" data-image-alt=\"Figure 3\" data-pmc-id=\"PMC4350901\" data-figure-id=\"Fig3\"> <img decoding=\"async\" id=\"article-image-2\" class=\"figure-thumb\" src=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig3_HTML.gif\" alt=\"Figure 3\" \/> <\/a><\/p>\n<div class=\"figure-caption figure-caption-text\" aria-hidden=\"true\">\n<div class=\"caption-wrap\">\n<p>Figure 3<\/p>\n<div class=\"figure-caption-contents\"><b> Designing primers based on defined\u2026 <\/b><\/div>\n<\/div>\n<\/div>\n<\/figure>\n<figure class=\"figure-item \" data-slide-index=\"3\" data-label-slug=\"figure-4\"><a class=\"figure-link\" role=\"button\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig4_HTML.jpg\" aria-describedby=\"figure-caption-3\" data-image-width=\"600\" data-image-height=\"497\" data-image-alt=\"Figure 4\" data-pmc-id=\"PMC4350901\" data-figure-id=\"Fig4\"> <img decoding=\"async\" id=\"article-image-3\" class=\"figure-thumb\" src=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig4_HTML.gif\" alt=\"Figure 4\" \/> <\/a><\/p>\n<div class=\"figure-caption figure-caption-text\" aria-hidden=\"true\">\n<div class=\"caption-wrap\">\n<p>Figure 4<\/p>\n<div class=\"figure-caption-contents\"><b> Sequence analysis of the PCR\u2026 <\/b><\/div>\n<\/div>\n<\/div>\n<\/figure>\n<figure class=\"figure-item \" data-slide-index=\"4\" data-label-slug=\"figure-5\"><a class=\"figure-link\" role=\"button\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig5_HTML.jpg\" aria-describedby=\"figure-caption-4\" data-image-width=\"600\" data-image-height=\"661\" data-image-alt=\"Figure 5\" data-pmc-id=\"PMC4350901\" data-figure-id=\"Fig5\"> <img decoding=\"async\" id=\"article-image-4\" class=\"figure-thumb\" src=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig5_HTML.gif\" alt=\"Figure 5\" \/> <\/a><\/p>\n<div class=\"figure-caption figure-caption-text\" aria-hidden=\"true\">\n<div class=\"caption-wrap\">\n<p>Figure 5<\/p>\n<div class=\"figure-caption-contents\"><b> Vector and insert plasmid maps\u2026 <\/b><\/div>\n<\/div>\n<\/div>\n<\/figure>\n<figure class=\"figure-item \" data-slide-index=\"5\" data-label-slug=\"figure-6\"><a class=\"figure-link\" role=\"button\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig6_HTML.jpg\" aria-describedby=\"figure-caption-5\" data-image-width=\"600\" data-image-height=\"198\" data-image-alt=\"Figure 6\" data-pmc-id=\"PMC4350901\" data-figure-id=\"Fig6\"> <img decoding=\"async\" id=\"article-image-5\" class=\"figure-thumb\" src=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/instance\/4350901\/bin\/13036_2014_161_Fig6_HTML.gif\" alt=\"Figure 6\" \/> <\/a><\/p>\n<div class=\"figure-caption figure-caption-text\" aria-hidden=\"true\">\n<div class=\"caption-wrap\">\n<p>Figure 6<\/p>\n<div class=\"figure-caption-contents\"><b> Screening of the final<\/b><\/div>\n<div>SOURCE:https:\/\/pubmed.ncbi.nlm.nih.gov\/25745516\/<\/div>\n<\/div>\n<div><\/div>\n<div>\u039c\u0395\u03a4\u0391\u03a6\u03a1\u0391\u03a3\u0397 \u03a4\u039f\u03a5 \u0391\u03a1\u0398\u03a1\u039f\u03a5:<\/div>\n<div>\n<p>&nbsp;<\/p>\n<p><strong><span style=\"font-size: 24pt;\">\u0397 \u03c7\u03c1\u03ae\u03c3\u03b7 \u03c4\u03bf\u03c5 PCR \u03c4\u03b5\u03c3\u03c4 \u03c9\u03c2 \u03b5\u03ba\u03c0\u03b1\u03b9\u03b4\u03b5\u03c5\u03c4\u03b9\u03ba\u03cc\u03c2 \u03bf\u03b4\u03b7\u03b3\u03cc\u03c2 \u03b3\u03b9\u03b1 \u03c4\u03b7\u03bd \u03ba\u03c5\u03c4\u03c4\u03b1\u03c1\u03b9\u03ba\u03ae \u03bc\u03b7\u03c7\u03b1\u03bd\u03b9\u03ba\u03ae<\/span><\/strong><\/p>\n<p><span style=\"font-size: 18pt;\"><strong>\u039c\u03bf\u03c1\u03b9\u03b1\u03ba\u03ae \u03ba\u03bb\u03c9\u03bd\u03bf\u03c0\u03bf\u03af\u03b7\u03c3\u03b7 \u03bc\u03b5 \u03c7\u03c1\u03ae\u03c3\u03b7 \u03b1\u03bb\u03c5\u03c3\u03b9\u03b4\u03c9\u03c4\u03ae\u03c2 \u03b1\u03bd\u03c4\u03af\u03b4\u03c1\u03b1\u03c3\u03b7\u03c2 \u03c0\u03bf\u03bb\u03c5\u03bc\u03b5\u03c1\u03ac\u03c3\u03b7\u03c2, \u03ad\u03bd\u03b1\u03c2 \u03b5\u03ba\u03c0\u03b1\u03b9\u03b4\u03b5\u03c5\u03c4\u03b9\u03ba\u03cc\u03c2 \u03bf\u03b4\u03b7\u03b3\u03cc\u03c2 \u03b3\u03b9\u03b1 \u03c4\u03b7\u03bd \u03ba\u03c5\u03c4\u03c4\u03b1\u03c1\u03b9\u03ba\u03ae \u03bc\u03b7\u03c7\u03b1\u03bd\u03b9\u03ba\u03ae<\/strong><\/span><\/p>\n<p>Sayed Shahabuddin Hoseini 1, Martin G Sauer 2<\/p>\n<p>\u0391\u03a1\u03a7\u0395\u0399\u0391<\/p>\n<p>PMID: 25745516<\/p>\n<p>PMCID: PMC4350901<\/p>\n<p>DOI: 10.1186\/1754-1611-9-2<\/p>\n<p>\u0399\u03c3\u03c4\u03bf\u03c1\u03b9\u03ba\u03cc: \u03a4\u03b9\u03c2 \u03c4\u03b5\u03bb\u03b5\u03c5\u03c4\u03b1\u03af\u03b5\u03c2 \u03b4\u03b5\u03ba\u03b1\u03b5\u03c4\u03af\u03b5\u03c2, \u03b7 \u03bc\u03bf\u03c1\u03b9\u03b1\u03ba\u03ae \u03ba\u03bb\u03c9\u03bd\u03bf\u03c0\u03bf\u03af\u03b7\u03c3\u03b7 \u03ad\u03c7\u03b5\u03b9 \u03bc\u03b5\u03c4\u03b1\u03bc\u03bf\u03c1\u03c6\u03ce\u03c3\u03b5\u03b9 \u03c4\u03b9\u03c2 \u03b2\u03b9\u03bf\u03bb\u03bf\u03b3\u03b9\u03ba\u03ad\u03c2 \u03b5\u03c0\u03b9\u03c3\u03c4\u03ae\u03bc\u03b5\u03c2. \u0388\u03c7\u03bf\u03bd\u03c4\u03b1\u03c2 \u03b5\u03c0\u03b7\u03c1\u03b5\u03ac\u03c3\u03b5\u03b9 \u03b2\u03b1\u03b8\u03b9\u03ac \u03b4\u03b9\u03ac\u03c6\u03bf\u03c1\u03bf\u03c5\u03c2 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\u03bc\u03b7\u03c7\u03b1\u03bd\u03b9\u03ba\u03ae\u03c2. \u0395\u03b4\u03ce, \u03b7 \u03b1\u03bb\u03c5\u03c3\u03b9\u03b4\u03c9\u03c4\u03ae \u03b1\u03bd\u03c4\u03af\u03b4\u03c1\u03b1\u03c3\u03b7 \u03c0\u03bf\u03bb\u03c5\u03bc\u03b5\u03c1\u03ac\u03c3\u03b7\u03c2 (PCR) \u03b1\u03bd\u03c4\u03b9\u03c0\u03c1\u03bf\u03c3\u03c9\u03c0\u03b5\u03cd\u03b5\u03b9 \u03ad\u03bd\u03b1 \u03b1\u03c0\u03cc \u03c4\u03b1 \u03c0\u03b9\u03bf \u03b2\u03b1\u03c3\u03b9\u03ba\u03ac \u03b5\u03c1\u03b3\u03b1\u03bb\u03b5\u03af\u03b1. \u039b\u03cc\u03b3\u03c9 \u03c4\u03b7\u03c2 \u03b5\u03bc\u03c6\u03ac\u03bd\u03b9\u03c3\u03b7\u03c2 \u03bd\u03ad\u03c9\u03bd \u03ba\u03b1\u03b9 \u03b1\u03c0\u03bf\u03c4\u03b5\u03bb\u03b5\u03c3\u03bc\u03b1\u03c4\u03b9\u03ba\u03ce\u03bd \u03b1\u03bd\u03c4\u03b9\u03b4\u03c1\u03b1\u03c3\u03c4\u03b7\u03c1\u03af\u03c9\u03bd PCR, \u03ba\u03b9\u03c4 \u03ba\u03bb\u03c9\u03bd\u03bf\u03c0\u03bf\u03af\u03b7\u03c3\u03b7\u03c2 \u03ba\u03b1\u03b9 \u03bb\u03bf\u03b3\u03b9\u03c3\u03bc\u03b9\u03ba\u03bf\u03cd, \u03c5\u03c0\u03ac\u03c1\u03c7\u03b5\u03b9 \u03b1\u03bd\u03ac\u03b3\u03ba\u03b7 \u03b3\u03b9\u03b1 \u03ad\u03bd\u03b1 \u03c3\u03c5\u03bd\u03bf\u03c0\u03c4\u03b9\u03ba\u03cc \u03ba\u03b1\u03b9 \u03bf\u03bb\u03bf\u03ba\u03bb\u03b7\u03c1\u03c9\u03bc\u03ad\u03bd\u03bf \u03c0\u03c1\u03c9\u03c4\u03cc\u03ba\u03bf\u03bb\u03bb\u03bf \u03c0\u03bf\u03c5 \u03bd\u03b1 \u03b5\u03be\u03b7\u03b3\u03b5\u03af \u03cc\u03bb\u03b1 \u03c4\u03b1 \u03c3\u03c4\u03ac\u03b4\u03b9\u03b1 \u03c4\u03b7\u03c2 \u03ba\u03bb\u03c9\u03bd\u03bf\u03c0\u03bf\u03af\u03b7\u03c3\u03b7\u03c2 PCR \u03be\u03b5\u03ba\u03b9\u03bd\u03ce\u03bd\u03c4\u03b1\u03c2 \u03b1\u03c0\u03cc \u03c4\u03bf \u03c3\u03c7\u03b5\u03b4\u03b9\u03b1\u03c3\u03bc\u03cc \u03c4\u03bf\u03c5 \u03b5\u03ba\u03ba\u03b9\u03bd\u03b7\u03c4\u03ae, \u03c4\u03b7\u03bd \u03b5\u03ba\u03c4\u03ad\u03bb\u03b5\u03c3\u03b7 PCR, \u03c4\u03bf\u03bd \u03c0\u03c1\u03bf\u03c3\u03b4\u03b9\u03bf\u03c1\u03b9\u03c3\u03bc\u03cc \u03b1\u03bb\u03bb\u03b7\u03bb\u03bf\u03c5\u03c7\u03af\u03b1\u03c2 \u03c0\u03c1\u03bf\u03ca\u03cc\u03bd\u03c4\u03c9\u03bd PCR, \u03c4\u03b7\u03bd \u03b1\u03bd\u03ac\u03bb\u03c5\u03c3\u03b7 \u03c4\u03c9\u03bd \u03b4\u03b5\u03b4\u03bf\u03bc\u03ad\u03bd\u03b1 \u03b1\u03bb\u03bb\u03b7\u03bb\u03bf\u03c5\u03c7\u03af\u03b1\u03c2 \u03ba\u03b1\u03b9 \u03c4\u03ad\u03bb\u03bf\u03c2 \u03b7 \u03b1\u03be\u03b9\u03bf\u03bb\u03cc\u03b3\u03b7\u03c3\u03b7 \u03c4\u03b7\u03c2 \u03b3\u03bf\u03bd\u03b9\u03b4\u03b9\u03b1\u03ba\u03ae\u03c2 \u03ad\u03ba\u03c6\u03c1\u03b1\u03c3\u03b7\u03c2. \u03a3\u03c4\u03cc\u03c7\u03bf\u03c2 \u03b1\u03c5\u03c4\u03bf\u03cd \u03c4\u03bf\u03c5 \u03b5\u03b3\u03b3\u03c1\u03ac\u03c6\u03bf\u03c5 \u03bc\u03b5\u03b8\u03bf\u03b4\u03bf\u03bb\u03bf\u03b3\u03af\u03b1\u03c2 \u03b5\u03af\u03bd\u03b1\u03b9 \u03bd\u03b1 \u03c0\u03b1\u03c1\u03ad\u03c7\u03b5\u03b9 \u03ad\u03bd\u03b1 \u03bf\u03bb\u03bf\u03ba\u03bb\u03b7\u03c1\u03c9\u03bc\u03ad\u03bd\u03bf \u03c0\u03c1\u03c9\u03c4\u03cc\u03ba\u03bf\u03bb\u03bb\u03bf \u03bc\u03b5 \u03ad\u03bd\u03b1 \u03b2\u03b9\u03ce\u03c3\u03b9\u03bc\u03bf \u03c0\u03b1\u03c1\u03ac\u03b4\u03b5\u03b9\u03b3\u03bc\u03b1 \u03b3\u03b9\u03b1 \u03c4\u03b7\u03bd \u03b5\u03c6\u03b1\u03c1\u03bc\u03bf\u03b3\u03ae \u03c4\u03b7\u03c2 PCR \u03c3\u03c4\u03b7\u03bd \u03ba\u03bb\u03c9\u03bd\u03bf\u03c0\u03bf\u03af\u03b7\u03c3\u03b7 \u03b3\u03bf\u03bd\u03b9\u03b4\u03af\u03c9\u03bd.<\/p>\n<p>\u0391\u03c0\u03bf\u03c4\u03b5\u03bb\u03ad\u03c3\u03bc\u03b1\u03c4\u03b1: \u03a0\u03b1\u03c1\u03b1\u03b4\u03b5\u03b9\u03b3\u03bc\u03b1\u03c4\u03b9\u03ba\u03ac, \u03b7 \u03b1\u03bb\u03bb\u03b7\u03bb\u03bf\u03c5\u03c7\u03af\u03b1 \u03c4\u03bf\u03c5 \u03c6\u03b8\u03bf\u03c1\u03af\u03b6\u03bf\u03bd\u03c4\u03bf\u03c2 \u03b3\u03bf\u03bd\u03b9\u03b4\u03af\u03bf\u03c5 tdTomato \u03b5\u03bd\u03b9\u03c3\u03c7\u03cd\u03b8\u03b7\u03ba\u03b5 \u03bc\u03b5 \u03b5\u03ba\u03ba\u03b9\u03bd\u03b7\u03c4\u03ad\u03c2 PCR \u03cc\u03c0\u03bf\u03c5 \u03b5\u03bd\u03c3\u03c9\u03bc\u03b1\u03c4\u03ce\u03b8\u03b7\u03ba\u03b1\u03bd \u03ba\u03b1\u03c4\u03ac\u03bb\u03bb\u03b7\u03bb\u03b5\u03c2 \u03b8\u03ad\u03c3\u03b5\u03b9\u03c2 \u03c0\u03b5\u03c1\u03b9\u03bf\u03c1\u03b9\u03c3\u03c4\u03b9\u03ba\u03ce\u03bd \u03b5\u03bd\u03b6\u03cd\u03bc\u03c9\u03bd. \u0395\u03c0\u03b5\u03be\u03b7\u03b3\u03bf\u03cd\u03bd\u03c4\u03b1\u03b9 \u03c4\u03b1 \u03c0\u03c1\u03b1\u03ba\u03c4\u03b9\u03ba\u03ac \u03ba\u03c1\u03b9\u03c4\u03ae\u03c1\u03b9\u03b1 \u03b3\u03b9\u03b1 \u03c4\u03b7\u03bd \u03b5\u03c0\u03b9\u03bb\u03bf\u03b3\u03ae \u03c4\u03c9\u03bd \u03c0\u03b5\u03c1\u03b9\u03bf\u03c1\u03b9\u03c3\u03c4\u03b9\u03ba\u03ce\u03bd \u03b5\u03bd\u03b6\u03cd\u03bc\u03c9\u03bd \u03ba\u03b1\u03b9 \u03bf \u03c3\u03c7\u03b5\u03b4\u03b9\u03b1\u03c3\u03bc\u03cc\u03c2 \u03c4\u03c9\u03bd \u03b5\u03ba\u03ba\u03b9\u03bd\u03b7\u03c4\u03ce\u03bd PCR. \u03a0\u03b1\u03c1\u03bf\u03c5\u03c3\u03b9\u03ac\u03b6\u03b5\u03c4\u03b1\u03b9 \u03b1\u03c0\u03bf\u03c4\u03b5\u03bb\u03b5\u03c3\u03bc\u03b1\u03c4\u03b9\u03ba\u03ae \u03ba\u03bb\u03c9\u03bd\u03bf\u03c0\u03bf\u03af\u03b7\u03c3\u03b7 \u03c0\u03c1\u03bf\u03ca\u03cc\u03bd\u03c4\u03c9\u03bd PCR \u03c3\u03b5 \u03ad\u03bd\u03b1 \u03c0\u03bb\u03b1\u03c3\u03bc\u03af\u03b4\u03b9\u03bf \u03b3\u03b9\u03b1 \u03c0\u03c1\u03bf\u03c3\u03b4\u03b9\u03bf\u03c1\u03b9\u03c3\u03bc\u03cc \u03b1\u03bb\u03bb\u03b7\u03bb\u03bf\u03c5\u03c7\u03af\u03b1\u03c2 \u03ba\u03b1\u03b9 \u03b4\u03c9\u03c1\u03b5\u03ac\u03bd \u03bb\u03bf\u03b3\u03b9\u03c3\u03bc\u03b9\u03ba\u03cc \u03b2\u03b1\u03c3\u03b9\u03c3\u03bc\u03ad\u03bd\u03bf \u03c3\u03c4\u03bf\u03bd \u03b9\u03c3\u03c4\u03cc \u03b3\u03b9\u03b1 \u03c4\u03b7 \u03b4\u03b9\u03b1\u03b4\u03bf\u03c7\u03b9\u03ba\u03ae \u03b1\u03bd\u03ac\u03bb\u03c5\u03c3\u03b7 \u03c4\u03c9\u03bd \u03b4\u03b5\u03b4\u03bf\u03bc\u03ad\u03bd\u03c9\u03bd \u03c0\u03c1\u03bf\u03c3\u03b4\u03b9\u03bf\u03c1\u03b9\u03c3\u03bc\u03bf\u03cd \u03b1\u03bb\u03bb\u03b7\u03bb\u03bf\u03c5\u03c7\u03af\u03b1\u03c2. \u03a4\u03ad\u03bb\u03bf\u03c2, \u03b7 \u03b5\u03c0\u03b9\u03b2\u03b5\u03b2\u03b1\u03af\u03c9\u03c3\u03b7 \u03c4\u03b7\u03c2 \u03b5\u03c0\u03b9\u03c4\u03c5\u03c7\u03bf\u03cd\u03c2 \u03ba\u03bb\u03c9\u03bd\u03bf\u03c0\u03bf\u03af\u03b7\u03c3\u03b7\u03c2 \u03b5\u03be\u03b7\u03b3\u03b5\u03af\u03c4\u03b1\u03b9 \u03c7\u03c1\u03b7\u03c3\u03b9\u03bc\u03bf\u03c0\u03bf\u03b9\u03ce\u03bd\u03c4\u03b1\u03c2 \u03ad\u03bd\u03b1 \u03c6\u03b8\u03bf\u03c1\u03af\u03b6\u03bf\u03bd \u03b3\u03bf\u03bd\u03af\u03b4\u03b9\u03bf \u03b5\u03bd\u03b4\u03b9\u03b1\u03c6\u03ad\u03c1\u03bf\u03bd\u03c4\u03bf\u03c2 \u03ba\u03b1\u03b9 \u03ba\u03cd\u03c4\u03c4\u03b1\u03c1\u03b1-\u03c3\u03c4\u03cc\u03c7\u03bf\u03c5\u03c2 \u03c0\u03bf\u03bd\u03c4\u03b9\u03ba\u03bf\u03cd.<\/p>\n<p>\u03a3\u03c5\u03bc\u03c0\u03b5\u03c1\u03ac\u03c3\u03bc\u03b1\u03c4\u03b1: \u03a7\u03c1\u03b7\u03c3\u03b9\u03bc\u03bf\u03c0\u03bf\u03b9\u03ce\u03bd\u03c4\u03b1\u03c2 \u03ad\u03bd\u03b1 \u03c0\u03c1\u03b1\u03ba\u03c4\u03b9\u03ba\u03cc \u03c0\u03b1\u03c1\u03ac\u03b4\u03b5\u03b9\u03b3\u03bc\u03b1, \u03b5\u03b9\u03c3\u03ae\u03c7\u03b8\u03b7 \u03ad\u03bd\u03b1 \u03bf\u03bb\u03bf\u03ba\u03bb\u03b7\u03c1\u03c9\u03bc\u03ad\u03bd\u03bf \u03c0\u03c1\u03c9\u03c4\u03cc\u03ba\u03bf\u03bb\u03bb\u03bf \u03b2\u03b1\u03c3\u03b9\u03c3\u03bc\u03ad\u03bd\u03bf \u03c3\u03b5 PCR \u03bc\u03b5 \u03c3\u03b7\u03bc\u03b1\u03bd\u03c4\u03b9\u03ba\u03ad\u03c2 \u03c3\u03c5\u03bc\u03b2\u03bf\u03c5\u03bb\u03ad\u03c2. \u0391\u03c5\u03c4\u03cc \u03c4\u03bf \u03ad\u03b3\u03b3\u03c1\u03b1\u03c6\u03bf \u03bc\u03b5\u03b8\u03bf\u03b4\u03bf\u03bb\u03bf\u03b3\u03af\u03b1\u03c2 \u03bc\u03c0\u03bf\u03c1\u03b5\u03af \u03bd\u03b1 \u03c7\u03c1\u03b7\u03c3\u03b9\u03bc\u03b5\u03cd\u03c3\u03b5\u03b9 \u03c9\u03c2 \u03bf\u03b4\u03b9\u03ba\u03cc\u03c2 \u03c7\u03ac\u03c1\u03c4\u03b7\u03c2 \u03b3\u03b9\u03b1 \u03b5\u03c1\u03b5\u03c5\u03bd\u03b7\u03c4\u03ad\u03c2 \u03c0\u03bf\u03c5 \u03b8\u03ad\u03bb\u03bf\u03c5\u03bd \u03bd\u03b1 \u03b5\u03ba\u03bc\u03b5\u03c4\u03b1\u03bb\u03bb\u03b5\u03c5\u03c4\u03bf\u03cd\u03bd \u03b3\u03c1\u03ae\u03b3\u03bf\u03c1\u03b1 \u03c4\u03b7 \u03b4\u03cd\u03bd\u03b1\u03bc\u03b7 \u03c4\u03b7\u03c2 PCR-\u03ba\u03bb\u03c9\u03bd\u03bf\u03c0\u03bf\u03af\u03b7\u03c3\u03b7\u03c2, \u03b1\u03bb\u03bb\u03ac \u03ad\u03c7\u03bf\u03c5\u03bd \u03c4\u03b7\u03bd \u03ba\u03cd\u03c1\u03b9\u03b1 \u03b5\u03c3\u03c4\u03af\u03b1\u03c3\u03ae \u03c4\u03bf\u03c5\u03c2 \u03c3\u03b5 \u03bb\u03b5\u03b9\u03c4\u03bf\u03c5\u03c1\u03b3\u03b9\u03ba\u03ad\u03c2 in vitro \u03ba\u03b1\u03b9 in vivo \u03c0\u03c4\u03c5\u03c7\u03ad\u03c2 \u03c4\u03b7\u03c2 \u03ba\u03c5\u03c4\u03c4\u03b1\u03c1\u03b9\u03ba\u03ae\u03c2 \u03bc\u03b7\u03c7\u03b1\u03bd\u03b9\u03ba\u03ae\u03c2.<\/p>\n<p>\u039b\u03ad\u03be\u03b5\u03b9\u03c2-\u03ba\u03bb\u03b5\u03b9\u03b4\u03b9\u03ac: \u0392\u03b9\u03bf\u03bb\u03bf\u03b3\u03b9\u03ba\u03ae \u03bc\u03b7\u03c7\u03b1\u03bd\u03b9\u03ba\u03ae; \u039a\u03bb\u03c9\u03bd\u03bf\u03c0\u03bf\u03af\u03b7\u03c3\u03b7; \u0395\u03ba\u03c0\u03b1\u03b9\u03b4\u03b5\u03c5\u03c4\u03b9\u03ba\u03cc\u03c2 \u03bf\u03b4\u03b7\u03b3\u03cc\u03c2; \u0391\u03bb\u03c5\u03c3\u03b9\u03b4\u03c9\u03c4\u03ae \u03b1\u03bd\u03c4\u03af\u03b4\u03c1\u03b1\u03c3\u03b7 \u03c0\u03bf\u03bb\u03c5\u03bc\u03b5\u03c1\u03ac\u03c3\u03b7\u03c2 (PCR); \u0391\u03bd\u03b1\u03c3\u03c5\u03bd\u03b4\u03c5\u03b1\u03c3\u03bc\u03ad\u03bd\u03bf DNA; \u039c\u03b5\u03c4\u03b1\u03c6\u03bf\u03c1\u03ac; \u039c\u03b5\u03c4\u03b1\u03bc\u03cc\u03bb\u03c5\u03bd\u03c3\u03b7.<\/p>\n<p>\u03a0\u0397\u0393\u0397:https:\/\/pubmed.ncbi.nlm.nih.gov\/25745516\/<\/p>\n<\/div>\n<\/div>\n<\/figure>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Molecular cloning using polymerase chain reaction, an educational guide for cellular engineering Sayed Shahabuddin Hoseini\u00a0 1 ,\u00a0Martin G Sauer\u00a0 2 Affiliations PMID: 25745516 PMCID: PMC4350901 DOI: 10.1186\/1754-1611-9-2 Free PMC article Abstract Background: Over the last decades, molecular cloning has transformed biological sciences. Having profoundly impacted various areas such as basic science, clinical, pharmaceutical, and environmental &hellip; <\/p>\n<p><a class=\"more-link btn\" href=\"https:\/\/evaggelatos.com\/?p=29345\">\u03a3\u03c5\u03bd\u03ad\u03c7\u03b5\u03b9\u03b1 \u03b1\u03bd\u03ac\u03b3\u03bd\u03c9\u03c3\u03b7\u03c2<\/a><\/p>\n","protected":false},"author":2,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[85],"tags":[217,90],"class_list":["post-29345","post","type-post","status-publish","format-standard","hentry","category-85","tag-217","tag-90","item-wrap"],"_links":{"self":[{"href":"https:\/\/evaggelatos.com\/index.php?rest_route=\/wp\/v2\/posts\/29345","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/evaggelatos.com\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/evaggelatos.com\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/evaggelatos.com\/index.php?rest_route=\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/evaggelatos.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=29345"}],"version-history":[{"count":5,"href":"https:\/\/evaggelatos.com\/index.php?rest_route=\/wp\/v2\/posts\/29345\/revisions"}],"predecessor-version":[{"id":29351,"href":"https:\/\/evaggelatos.com\/index.php?rest_route=\/wp\/v2\/posts\/29345\/revisions\/29351"}],"wp:attachment":[{"href":"https:\/\/evaggelatos.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=29345"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/evaggelatos.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=29345"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/evaggelatos.com\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=29345"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}