validation of a genus-specific gene; tps, used as internal control in quantitative real time pcr of transgenic cotton

نویسندگان

seyde mehri javadi

masuod tohidfar

ali haghnazari

shahin negari

چکیده

identification of genes with invariant levels of gene expression is a prerequisite for validating transcriptomicchanges accompanying development. ideally expression of these genes should be independent of themorphogenetic process or environmental condition.we report here the validation of internal control genei.e.tps (trehalose 6-phosphate-synthase) in cotton (gossypium spp), using taqman system in quantitativereal time pcr (qrt-pcr). the gene expression was tested in five different g. hirsutum cultivars includingcoker 312, acala sj, zeta 2, taghva, neishabour and a diploid wild type; g. barbadense. identicalamplicons were obtained within these cultivars. no amplifications was achieved when dna samples frombarley (hordeum vulgare), maize (zea mays), rice (oryza sativa), fig (ficus carica), pistachio (pistaciavera), yew (taxus baccata), wheat (tirticum aestivum), rose (rosa hybrida) and soybean (glycine max) wereused as template. therefore, it was confirmed that the primers and probes designed in this study were specificfor the identification and quantification of internal control gene. these results reveal the possibility of usingthe tps gene as an internal control in cotton. in another word, this gene would be a suitable candidate as areference gene in examination of gene expression, detection of transgenic cotton and determining e thezygosity as well as the copy number of the transgene.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Validation of a genus-specific gene; TPS, used as internal control in quantitative Real Time PCR of transgenic cotton

Identification of genes with invariant levels of gene expression is a prerequisite for validating transcriptomic changes accompanying development. Ideally expression of these genes should be independent of the morphogenetic process or environmental condition.We report here the validation of internal control gene i.e.TPS (trehalose 6-phosphate-synthase) in cotton (Gossypium spp), using TaqMan sy...

متن کامل

Quality Control of Widely Used Therapeutic Recombinant Proteins by a Novel Real-Time PCR Approac

Background: Existence of bacterial host cell DNA contamination in biopharmaceutical products is a potential risk factor for patients receiving these drugs. Hence, the quantity of contamination must be controlled under the regulatory standards. Although different methods such as hybridization assays have been employed to determine DNA impurities, these methods are labor, intensive and rather exp...

متن کامل

Validation of a quantitative method for real time PCR kinetics.

Real time RT-PCR is the most sensitive method for quantitation of gene expression levels. The accuracy can be dependent on the mathematical model on which the quantitative methods are based. The generally accepted mathematical model assumes that amplification efficiencies are equal at the exponential phase of the reactions for the same amplicon. However, no methods are available to test the ass...

متن کامل

Quantitative Real-Time PCR

Unlike classical end-point analysis PCR, real-time PCR provides the data required for quantification of the target nucleic acid. The results can be expressed in absolute terms by reference to external quantified standards or in relative terms compared to another target sequence present within the sample. Absolute quantification requires that the efficiency of the amplification reaction is the s...

متن کامل

Real time quantitative PCR.

We have developed a novel "real time" quantitative PCR method. The method measures PCR product accumulation through a dual-labeled fluorogenic probe (i.e., TaqMan Probe). This method provides very accurate and reproducible quantitation of gene copies. Unlike other quantitative PCR methods, real-time PCR does not require post-PCR sample handling, preventing potential PCR product carry-over conta...

متن کامل

Real-time quantitative PCR.

Those who have worked in the field of quantitative polymerase chain reaction (PCR) since the early 1990s have accepted many of the tedious aspects of the early assays as routine. Difficulties associated with early quantitative PCR techniques included: (i) ensuring that the PCR was within the linear range of amplification (that portion where the PCR signal is directly proportional to the input c...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
journal of plant molecular breeding

ناشر: genetics and agricultural biotechnology institute of tabarestan (gabit)

ISSN 2322-3332

دوره 1

شماره 2 2013

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023