Establishment and in-house validation of stem-loop rt pcr method for microrna398 expression analysis
2015
Autori
Timotijević, GordanaMilisavljević, Mira
Nikolic, Dragana B.
Milovanović, Bosko M.
Nikolic, Dragana S.
Nikolic, Miroslav
Samardžić, Jelena T.
Članak u časopisu (Objavljena verzija)
Metapodaci
Prikaz svih podataka o dokumentuApstrakt
MicroRNAs (miRNAs) belong to the class of small non-coding RNAs which have important roles throughout development as well as in plant response to diverse environmental stresses. Some of plant miRNAs are essential for regulation and maintenance of nutritive homeostasis when nutrients are in excess or shortage comparing to optimal concentration for certain plant species. Better understanding of miRNAs functions implies development of efficient technology for profiling their gene expression. We set out to establish validate the methodology for miRNA gene expression analysis in cucumber grown under suboptimal mineral nutrient regimes, including iron deficiency. Reverse transcription by "stem-loop" primers in combination with Real time PCR method is one of potential approaches for quantification of miRNA gene expression. In this paper we presented a method for "stem loop" primer design specific for miR398, as well as reaction optimization and determination of Real time PCR efficiency. Provi...ng the accuracy of this method was imperative as "stem loop" RT which consider separate transcription of target and endogenous control. The method was verified by comparison of the obtained data with results of miR398 expression achieved using a commercial kit based on simultaneous conversion of all RNAs in cDNAs.
Ključne reči:
Real time PCR / miRNA gene expression analysis / iron deficiencyIzvor:
Genetika-Belgrade, 2015, 47, 2, 405-416Izdavač:
- Društvo genetičara Srbije, Beograd
Finansiranje / projekti:
- Molekularni mehanizmi odgovora biljaka na abiotički stres-uloga transkripcionih faktora i malih RNK i analiza genetičkog diverziteta biljnih kultura od interesa za poljoprivredu i biotehnologiju (RS-173005)
- Mineralni stres i adaptacije biljaka na marginalnim poljoprivrednim zemljištima (RS-173028)
DOI: 10.2298/GENSR1502405T
ISSN: 0534-0012
WoS: 000363055800004
Scopus: 2-s2.0-84945961001
Institucija/grupa
Institut za multidisciplinarna istraživanjaTY - JOUR AU - Timotijević, Gordana AU - Milisavljević, Mira AU - Nikolic, Dragana B. AU - Milovanović, Bosko M. AU - Nikolic, Dragana S. AU - Nikolic, Miroslav AU - Samardžić, Jelena T. PY - 2015 UR - http://rimsi.imsi.bg.ac.rs/handle/123456789/848 AB - MicroRNAs (miRNAs) belong to the class of small non-coding RNAs which have important roles throughout development as well as in plant response to diverse environmental stresses. Some of plant miRNAs are essential for regulation and maintenance of nutritive homeostasis when nutrients are in excess or shortage comparing to optimal concentration for certain plant species. Better understanding of miRNAs functions implies development of efficient technology for profiling their gene expression. We set out to establish validate the methodology for miRNA gene expression analysis in cucumber grown under suboptimal mineral nutrient regimes, including iron deficiency. Reverse transcription by "stem-loop" primers in combination with Real time PCR method is one of potential approaches for quantification of miRNA gene expression. In this paper we presented a method for "stem loop" primer design specific for miR398, as well as reaction optimization and determination of Real time PCR efficiency. Proving the accuracy of this method was imperative as "stem loop" RT which consider separate transcription of target and endogenous control. The method was verified by comparison of the obtained data with results of miR398 expression achieved using a commercial kit based on simultaneous conversion of all RNAs in cDNAs. PB - Društvo genetičara Srbije, Beograd T2 - Genetika-Belgrade T1 - Establishment and in-house validation of stem-loop rt pcr method for microrna398 expression analysis EP - 416 IS - 2 SP - 405 VL - 47 DO - 10.2298/GENSR1502405T ER -
@article{ author = "Timotijević, Gordana and Milisavljević, Mira and Nikolic, Dragana B. and Milovanović, Bosko M. and Nikolic, Dragana S. and Nikolic, Miroslav and Samardžić, Jelena T.", year = "2015", abstract = "MicroRNAs (miRNAs) belong to the class of small non-coding RNAs which have important roles throughout development as well as in plant response to diverse environmental stresses. Some of plant miRNAs are essential for regulation and maintenance of nutritive homeostasis when nutrients are in excess or shortage comparing to optimal concentration for certain plant species. Better understanding of miRNAs functions implies development of efficient technology for profiling their gene expression. We set out to establish validate the methodology for miRNA gene expression analysis in cucumber grown under suboptimal mineral nutrient regimes, including iron deficiency. Reverse transcription by "stem-loop" primers in combination with Real time PCR method is one of potential approaches for quantification of miRNA gene expression. In this paper we presented a method for "stem loop" primer design specific for miR398, as well as reaction optimization and determination of Real time PCR efficiency. Proving the accuracy of this method was imperative as "stem loop" RT which consider separate transcription of target and endogenous control. The method was verified by comparison of the obtained data with results of miR398 expression achieved using a commercial kit based on simultaneous conversion of all RNAs in cDNAs.", publisher = "Društvo genetičara Srbije, Beograd", journal = "Genetika-Belgrade", title = "Establishment and in-house validation of stem-loop rt pcr method for microrna398 expression analysis", pages = "416-405", number = "2", volume = "47", doi = "10.2298/GENSR1502405T" }
Timotijević, G., Milisavljević, M., Nikolic, D. B., Milovanović, B. M., Nikolic, D. S., Nikolic, M.,& Samardžić, J. T.. (2015). Establishment and in-house validation of stem-loop rt pcr method for microrna398 expression analysis. in Genetika-Belgrade Društvo genetičara Srbije, Beograd., 47(2), 405-416. https://doi.org/10.2298/GENSR1502405T
Timotijević G, Milisavljević M, Nikolic DB, Milovanović BM, Nikolic DS, Nikolic M, Samardžić JT. Establishment and in-house validation of stem-loop rt pcr method for microrna398 expression analysis. in Genetika-Belgrade. 2015;47(2):405-416. doi:10.2298/GENSR1502405T .
Timotijević, Gordana, Milisavljević, Mira, Nikolic, Dragana B., Milovanović, Bosko M., Nikolic, Dragana S., Nikolic, Miroslav, Samardžić, Jelena T., "Establishment and in-house validation of stem-loop rt pcr method for microrna398 expression analysis" in Genetika-Belgrade, 47, no. 2 (2015):405-416, https://doi.org/10.2298/GENSR1502405T . .