کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5589899 | 1569838 | 2017 | 21 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Ct shift: A novel and accurate real-time PCR quantification model for direct comparison of different nucleic acid sequences and its application for transposon quantifications
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کلمات کلیدی
Cockayne syndrome BeGFPcomparative CtHSP90RT-PCRΔΔCtCSBqPCRQuantitative PCR - PCR کمیcoding sequence - توالی کدگذاریreverse transcription PCR - رونویسی معکوس PCRSleeping Beauty - زیبای خفتهCockayne syndrome - سکته CockayneCopy number - شماره کپی کنیدPromoter activity - فعالیت تبلیغاتیrelative quantity - مقدار نسبیsplice variant - نوع اتصالاتquantitative polymerase chain reaction - واکنش زنجیره ای پلیمراز کمیHeat shock protein 90 - پروتئین شوک حرارت 90enhanced green fluorescent protein - پروتئین فلورسنت سبز افزایش یافته استthreshold cycle - چرخه آستانهCdS - کادمیم سولفید، سولفید کادمیم
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
There are numerous applications of quantitative PCR for both diagnostic and basic research. As in many other techniques the basis of quantification is that comparisons are made between different (unknown and known or reference) specimens of the same entity. When the aim is to compare real quantities of different species in samples, one cannot escape their separate precise absolute quantification. We have established a simple and reliable method for this purpose (Ct shift method) which combines the absolute and the relative approach. It requires a plasmid standard containing both sequences of amplicons to be compared (e.g. the target of interest and the endogenous control). It can serve as a reference sample with equal copies of templates for both targets. Using the ÎÎCt formula we can quantify the exact ratio of the two templates in each unknown sample. The Ct shift method has been successfully applied for transposon gene copy measurements, as well as for comparison of different mRNAs in cDNA samples. This study provides the proof of concept and introduces some potential applications of the method; the absolute nature of results even without the need for real reference samples can contribute to the universality of the method and comparability of different studies.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Gene - Volume 598, 20 January 2017, Pages 43-49
Journal: Gene - Volume 598, 20 January 2017, Pages 43-49
نویسندگان
Orsolya Kolacsek, EnikÅ Pergel, Nóra Varga, Ágota Apáti, Tamás I. Orbán,