{"id":363,"date":"2021-09-19T12:36:16","date_gmt":"2021-09-19T12:36:16","guid":{"rendered":"http:\/\/longgene.com.tr\/?p=363"},"modified":"2021-10-10T14:46:30","modified_gmt":"2021-10-10T14:46:30","slug":"rt-pcr-result-lower-amplification","status":"publish","type":"post","link":"https:\/\/longgene.com.tr\/?p=363","title":{"rendered":"RT-PCR Result: Lower Amplification"},"content":{"rendered":"\n<figure class=\"wp-block-image size-full\"><img loading=\"lazy\" width=\"918\" height=\"662\" src=\"http:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/plot_low_amplification.png\" alt=\"\" class=\"wp-image-355\" srcset=\"https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/plot_low_amplification.png 918w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/plot_low_amplification-300x216.png 300w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/plot_low_amplification-768x554.png 768w\" sizes=\"(max-width: 918px) 100vw, 918px\" \/><\/figure>\n\n\n\n<p><strong>Low amplification plots may be caused by; <\/strong><\/p>\n\n\n\n<ul><li><strong>Low quality PCR strip tubes<\/strong>, optical tubes, plates or sealing may be used <\/li><li>Master mix may be melted <strong>many times<\/strong> (more than 3 times), efficiency may be down<\/li><li>Kits may be out of date, please check the <strong>expiration dates<\/strong> of the RNA isolation solutions &amp; diagnostic kits<\/li><li>There is a <strong>low <\/strong>amplification plots above, if there is no amplification (if there is only a single straight line on the screen) then you can perform the instrument &#8216;self test&#8217; function. <\/li><\/ul>\n\n\n\n<p><strong>Optimal Software &#8216;X&#8217; (Cycle axis) and &#8216;Y&#8217; (Fluorescent axis) scale adjustment may be a solution to see the amplification plots higher;<\/strong><\/p>\n\n\n\n<figure class=\"wp-block-image size-large\"><img loading=\"lazy\" width=\"1024\" height=\"616\" src=\"https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L1_Son-1024x616.png\" alt=\"pcr, qpcr, rt pcr, qrt pcr, real time, real time per, thermal cycler, longgene, longGene.com.tr, ivd ce, biotechnology, LongGene europe, europe pcr, disgnostics, snp, absolute quantification, relative quantification, dna, cdna, thermal protocol, LongGene, long gene, end point pcr, covid19, fast pcr, polymerase chain reaction, Q1000, Q1000+, optimal, longgene turkey, LongGene europe, pcr mix, rt-pcr mix, how to prepare pcr mix, 16 well rt pcr, optimal software\" class=\"wp-image-649\" srcset=\"https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L1_Son-1024x616.png 1024w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L1_Son-300x180.png 300w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L1_Son-768x462.png 768w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L1_Son-1536x923.png 1536w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L1_Son-1000x601.png 1000w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L1_Son-932x560.png 932w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L1_Son-600x361.png 600w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L1_Son.png 1760w\" sizes=\"(max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n\n\n\n<ul><li>Select &#8216;<strong>y<\/strong>&#8216; axis (Fluorescent value axis, when you select it correctly then the color changes to &#8216;<strong>blue<\/strong>&#8216;<\/li><li>Then use your <strong>mouse wheel<\/strong> to increase\/decrease the scale of the &#8216;y&#8217; axis. <\/li><li>Amplification curves become more clear to interpret the results.<\/li><\/ul>\n\n\n\n<hr class=\"wp-block-separator is-style-wide\"\/>\n\n\n\n<figure class=\"wp-block-image size-large\"><img loading=\"lazy\" width=\"1024\" height=\"541\" src=\"https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L2_Son-1024x541.png\" alt=\"pcr, qpcr, rt pcr, qrt pcr, real time, real time per, thermal cycler, longgene, longGene.com.tr, ivd ce, biotechnology, LongGene europe, europe pcr, disgnostics, snp, absolute quantification, relative quantification, dna, cdna, thermal protocol, LongGene, long gene, end point pcr, covid19, fast pcr, polymerase chain reaction, Q1000, Q1000+, optimal, longgene turkey, LongGene europe, pcr mix, rt-pcr mix, how to prepare pcr mix, 16 well rt pcr, optimal software\" class=\"wp-image-648\" srcset=\"https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L2_Son-1024x541.png 1024w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L2_Son-300x158.png 300w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L2_Son-768x406.png 768w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L2_Son-1536x812.png 1536w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L2_Son-1000x528.png 1000w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L2_Son-1060x560.png 1060w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L2_Son-600x317.png 600w, https:\/\/longgene.com.tr\/wp-content\/uploads\/2021\/09\/L2_Son.png 1730w\" sizes=\"(max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n\n\n\n<p>This will help you to check whether there is a sigmoidal application curve, or not. As above example, it&#8217;s clear that all internal controls are amplified as expected and there is no need to repeat any sample for this run. <\/p>\n\n\n\n<hr class=\"wp-block-separator is-style-wide\"\/>\n\n\n\n<p><strong>Related links;<\/strong><\/p>\n\n\n\n<div class=\"wp-block-buttons\">\n<div class=\"wp-block-button has-custom-font-size has-small-font-size\"><a class=\"wp-block-button__link\" href=\"http:\/\/longgene.com.tr\/?page_id=211\">96 WELL Real t\u0131me PCR<\/a><\/div>\n\n\n\n<div class=\"wp-block-button has-custom-font-size has-small-font-size\"><a class=\"wp-block-button__link\" href=\"http:\/\/longgene.com.tr\/?page_id=222\">48 well Real t\u0131me PCR<\/a><\/div>\n\n\n\n<div class=\"wp-block-button has-custom-font-size has-small-font-size\"><a class=\"wp-block-button__link\" href=\"http:\/\/longgene.com.tr\/?page_id=227\">16 well real t\u0131me pcr<\/a><\/div>\n\n\n\n<div class=\"wp-block-button has-custom-font-size has-small-font-size\"><a class=\"wp-block-button__link\" href=\"http:\/\/longgene.com.tr\/?page_id=204\">pcr d\u0131sposables<\/a><\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Low amplification plots may be caused by; Low quality PCR strip tubes, optical tubes, plates or sealing may be used Master mix may be melted many times (more than 3 times), efficiency may be down Kits may be out of date, please check the expiration dates of the RNA isolation [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[5],"tags":[9,10,12,13],"_links":{"self":[{"href":"https:\/\/longgene.com.tr\/index.php?rest_route=\/wp\/v2\/posts\/363"}],"collection":[{"href":"https:\/\/longgene.com.tr\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/longgene.com.tr\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/longgene.com.tr\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/longgene.com.tr\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=363"}],"version-history":[{"count":13,"href":"https:\/\/longgene.com.tr\/index.php?rest_route=\/wp\/v2\/posts\/363\/revisions"}],"predecessor-version":[{"id":660,"href":"https:\/\/longgene.com.tr\/index.php?rest_route=\/wp\/v2\/posts\/363\/revisions\/660"}],"wp:attachment":[{"href":"https:\/\/longgene.com.tr\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=363"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/longgene.com.tr\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=363"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/longgene.com.tr\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=363"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}