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Kantitatif RT-PCR (RT-qPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi ve Validasyonu

Yıl 2020, Cilt: 17 Sayı: 1, 76 - 83, 23.03.2020

Özgecan Korkmaz Ağaoğlu Özge Sidekli

https://doi.org/10.32707/ercivet.655015
Cited By: 1

Öz

Gen ekspresyon çalışmalarında farklı deneysel veya klinik koşullar altında genlerin ekspresyon seviyelerinin karşılaştırılması için kullanılan en yaygın teknik Kantitatif Gerçek Zamanlı Polimeraz Zincir Reaksiyonu (RT-qPCR)’dur. RT-qPCR’da güvenilir sonuçlar elde etmek için verilerin housekeeping genler (HKG) ile normalizasyonunun yapılması gerekmektedir. Bununla birlikte; çalışma tasarımına uygun en stabil HKG’nin kullanılması, çalışma sonuçlarının güvenilirliğini artırmaktadır. Güvenilir veri elde etmek için çalışma tasarımına uygun stabil HKG’nin seçimi ve validasyonu önemli bir adımdır ki bu ekspresyon bazlı çalışmalarda bütünleyici bir adım olmuştur. HKG seçimi için farklı yazılım programları geliştirilmiştir. Normalizasyon için seçilecek en stabil HKG’nin seçilmeden önce deneysel çalışmalar ile dikkatli bir şekilde değerlendirmesinin yapılması gerekmektedir. Bu derlemede; Kantitatif Gerçek Zamanlı PCR Deneylerinin Yayınlanması için Asgari Bilgi (MIQE), HKG’lerin genel özellikleri, RT-qPCR çalışmalarında kullanımları ile çalışma dizaynına göre en uygun ve stabil olan HKG’lerin seçimi ve validasyonu hakkında bilgiler özetlenmiştir.

Anahtar Kelimeler

Gen ekspresyonu housekeeping gen kantitatif RT-PCR referans gen

Kaynakça

  • Agaoglu AR, Agaoglu OK, Aslan S, Kocamuftuoglu M, Koker A, Cetin Y, Gungor O, Ozturk D, Saatcı M. The effects of presynch-ovsynch applications on endometrial Toll and Nod like receptor gene expressions in cows with repeat breeder syndrome (RBS). Turkish Society of Veterinary Gynaecology VII. National-I. International Congress, October, 15-18, 2017; Marmaris-Türkiye.
  • Andersen CL, Jensen JL, Ørntoft TF. Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 2004; 64(15): 5245-50.
  • Bär M, Bär D, Lehmann B. Selection and validation of candidate housekeeping genes for studies of human keratinocytes-review and recommendations. Jidonline 2009; 129(3): 535-7.
  • Barber RD, Harmer DW, Coleman RA, Clark BJ. GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissue. Physiol Genomics 2005; 21(3): 389-95.
  • Becker C, Hammerle-Fickinger A, Riedmaier I, Pfaffl MW. mRNA and microRNA quality control for RT-qPCR analysis. Methods 2010; 50(4):237-43.
  • Blomberg J, Andersson M, Faldt R. Differential pattern of oncogene and beta-actin expression in leukaemic cells from AML patients. J Haematol 1987; 65(1): 83-6.
  • Bogaert L, Van Poucke M, De Baere C, Peelman L, Gasthuys F, Martens A. Selection of a set of reliable reference genes for quantitative real-time PCR in normal equine skin and in equine sarcoids. BMC Biotechnol 2006; 6(24): 1-7.
  • Burns MJ, Nixon GJ, Foy CA, Harris N. Standardisation of data from real-time quantitative PCR methods – evaluation of outliers and comparison of calibration curves. BMC Biotechnol 2005; 5(31): 1-13.
  • Bustin SA, Beaulieu JF, Huggett J, Jaggi R, Kibenge FSB, Olsvik PA, Penning LC, Toegel S. MIQE precis: Practical implementation of minimum standard quidelines for fluorescence based quantitative real-time PCR expreriments. BMC Mol Biol 2010; 11(74): 1-5.
  • Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R,T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT. The MIQE Guidelines: Minimum information for publication of quantitive real-time PCR experiments. Clin Chem 2009; 55(4): 611-22.
  • Bustin SA. Why the need for qPCR publication guidelines?-The case for MIQE. Methods 2010b; 50(4): 217-26.
  • Chervoneva I, Li Y, Schulz S, Croker S, Wilson C, Waldman SA, Hyslop T. Selection of optimal reference genes for normalization in quantitative RT-PCR. BMC Bioinforma 2010; 11(253): 1-15.
  • Cinar MU, Islam MA, Uddin MJ, Tholen E, Tesfaye D, Looft C, Schellander K. Evaluation of suitable reference genes for gene expression studies in porcine alveolar macrophages in response to LPS and LTA. BMC Res Notes 2012; 5(107): 1-14.
  • Cross NC, Lin F, Goldman JM. Appropriate controls for reverse transcription polymerase chain reaction (RT-PCR). Br J Haematol 1994; 87(1): 218.
  • De Spiengelaere W, Dern-Wieloch J, Weigel R, Schumacher V, Schorle H, Nettersheim D, Bergmann M, Brehm R, Kliesch S, Vandekerckhove L, Fink C. Reference gene validation for RT-PCR, a note on different available software packages. PLoS One 2015; 10(3): e122515.
  • Du M, Wang YW, Yue PY, Zhou PY, Yao W, Li X, Ding XB, Liu XF, Guo H, Ma WZ. Selection of reference genes in canine uterine tissues. Genet Mol Res 2016; 15(2): 1-14. Eisenberg E, Levanon EY. Human housekeeping genes are compact. Trends Genet 2003; 19(7): 362-5.
  • Farre D, Bellora N, Mularoni L, Messeguer X, Alba MM. Housekeeping genes tend to Show reduced upstream sequence conservation. Genome Bio 2007; 8(7): RI40. Fleige S, Pfaffl MW. RNA integrity and the effect on the real-time qRT-PCR performance. Mol Aspects Med 2006; 27(2-3): 126-39.
  • Goldman M. Housekeeping Gene. Brenner S. Miller JH. eds. In: Encyclopedia of Genetics. Academic Press. 2001; pp. 978.
  • Gong S, Schmotzer CL, Zhou L. Evaluation of quantitative real-time PCR as a hepatitis C virus supplementary test after RIBA discontinuation. J Clin Lab Anal 2016; 30(5): 418-23.
  • Hashemipetroudi SH, Nematzadeh G, Ahmadian G, Yamchi A, Kuhlmann M. Assesment of DNA contamination in RNA samples based on ribosomal DNA. J Vis Exp 2018; 131(22): e55451.
  • Hayashi R. Gene expression and the impact of antioxidant supplements in the cataractous lens. Preedy VR. eds. Handbook of Nutrition, Diet and the Eye. London: Academic press 2014; p. 517-24.
  • Huggett J, Dheda K, Bustin S, Zumla A. Real-Time RT-PCR normalisation; strategies and considerations. Genes Immun 2005; 6(4): 279-84.
  • Jarczak J, Kaba J, Bagnicka E. The validation of housekeeping genes as a reference in quantitative Real Time PCR analysis: application in the milk somatic cells and frozen whole blood of goats infected with caprine arthritis encephalitis virüs. Gene 2014; 549(2): 280-5.
  • Kaur R, Sodhi M, Sharma A, Sharmi VL, Verma P, Swami SK, Kumari P, Mukesh M. Selection of suitable reference genes for normalizastion of quantitative RT-PCR expression data across twelve tissues of riverine buffaloes. PLoS One 2018; 13(3): E01191668.
  • Klatte M, Bauer P. Accurate Real-time reverse transcription quantitative PCR. Methods Mol Biol 2009; 479(4): 61-77.
  • Korkmaz Agaoglu O, Agaoglu AR, Ozmen O, Saatcı M, Schäfer-Somi S, Aslan S. Investigation of Insulin-like Growth Factor Gene Family in Cat Uterus During Pregnancy at mRNA and Protein Level. XXth International Congress of the European Veterinary Society for Small Animal Reproduction (EVSSAR), June 29-July 1, 2017; Vienna-Austria.
  • Kozera B, Rapacz M. Reference genes in real-time PCR. J Appl Genetics 2013; 54(4): 391-406.
  • Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitive PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25(4): 402-8.
  • Mahoney DJ, Carey K, Fu MH, Snow R, Cameron-Smith D, Parise G, Tarnopolsky MA. Real-time RT-PCR analysis of housekeeping genes in human skeletal muscle following acute exercise. 2004; 18(2): 226-31.
  • Nolan T, Bustin S. Procedures for Quality Control of RNA Samples for Use in Quantitative Reverse Transcription PCR. Keen JT Birch L. eds. In: Essentials of nucleic acid analysis: a robust approach. London: The royal society of Chemistry, 2008; pp. 189-207.
  • Panina Y, Germond A, Masui S, Watanabe TM. Validation of common housekeeping genes as reference for qPCR gene expression analysis during IPS reprogramming process. Sci Rep 2018; 8(1): 8716.
  • Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 2002; 30(9): e36.
  • Pfaffl MW, Tichopad A, Prgomet C, Neuvians TP. Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper-Excel-based tool using pair-wise correlations. Biotech Lett 2004; 26(6): 509-15.
  • Pfaffl MW. A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res 2001; 29(9): e45.
  • Sadek KH, Cagampang FR, Bruce KD, Shreeve N, Macklon N, Cheong Y. Variation in stability of housekeeping genes in endometrium of healthy and polycystic ovarian syndrome women. Hum Reprod 2012; 27(1): 251-6.
  • Segundo-Val IS, Sanz-Lozano CS. Introduction to the gene expression analysis. Methods Mol Biol 2016; 1434(3): 29-43.
  • Sikand K, Singh J, Ebron JS, Shukla GC. Housekeeping gene selection advisory: Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) and β-Actin are targets of miR-644a. PLoS One 2012; 7(10): e47510.
  • Ståhlberg A, Hakansson J, Xian X, Semb H, Kubista M. Properties of the reverse transcription reaction in mRNA quantification. Clin Chem 2004; 50(3): 509-15.
  • Sullivan-Gunn M, Hinch E, Vaughan V, Lewandowski P. Choosing a stable housekeeping gene and protein is essential in generating valid gene and protein expression results. Br J Cancer 2011; 104(6): 1055.
  • Tilli TM, Castro CS, Tuszynski JA, Carels N. A strategy to identify housekeeping genes suitable for analysis in breast cancer diseases. BMC Genomics 2016; 17(1): 639.
  • Uddin M, Cinar M, Tesfaye D, Looft C, Tholen E, Schellander K. Age-related changes in relative expression stability of commonly used housekeeping genes in selected porcine tissues. BMC Res Notes. 2011; 4(441): 1-13.
  • Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002; 3(7): 1-12.
  • VanGuilder HD, Vrana KE, Freeman WM. Twenty-five years of quantitative PCR for gene expression analysis. Biotechniques 2008; 44(5): 619-26.
  • Xie F, Sun G, Stiller JW and Zhang B. Genome-wide functional analysis of the cotton transcriptome by creating an integrated EST database. PLoS One 2011; 6(11): e26980.
  • Zhong H, Simons JW. Direct comparison of GAPDH, beta-actin, cyclophilin, and 28S rRNA as internal standards for quantifying RNA levels under hypoxia. Biochem Biophys Res Commun 1999; 259(3): 523-6.

Selection and Validation of Suitable Housekeeping Genes (HKGs) in Quantitative RT-PCR (RT-qPCR) Studies

Yıl 2020, Cilt: 17 Sayı: 1, 76 - 83, 23.03.2020

Özgecan Korkmaz Ağaoğlu Özge Sidekli

https://doi.org/10.32707/ercivet.655015
Cited By: 1

Öz

The quantitative Real-time Polymerase Chain Reaction (RT-qPCR) is the most common technique used to compare the expression levels of genes under different experimental or clinical conditions. Data should be normalized with housekeeping genes (HKGs) to obtain reliable results in RT-qPCR. Likewise, the use the most stable and suitable HKG for the study design improves the reliability of the study results. The selection and validation of a stable HKG according to the study design to obtain reliable data is an essential and integral step in expression-based studies. The several software programs have been developed for HKG selection. The most stable HKG to be selected for normalization needs to be carefully evaluated by experimental studies before selection. In this review, summarized Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE), characteristics of HKGs, selection and validation of most suitable and stable HKGs according to study design and, usage in RT-qPCR.

Anahtar Kelimeler

Gene expression housekeeping gene quantitative RT-PCR reference gene

Kaynakça

  • Agaoglu AR, Agaoglu OK, Aslan S, Kocamuftuoglu M, Koker A, Cetin Y, Gungor O, Ozturk D, Saatcı M. The effects of presynch-ovsynch applications on endometrial Toll and Nod like receptor gene expressions in cows with repeat breeder syndrome (RBS). Turkish Society of Veterinary Gynaecology VII. National-I. International Congress, October, 15-18, 2017; Marmaris-Türkiye.
  • Andersen CL, Jensen JL, Ørntoft TF. Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 2004; 64(15): 5245-50.
  • Bär M, Bär D, Lehmann B. Selection and validation of candidate housekeeping genes for studies of human keratinocytes-review and recommendations. Jidonline 2009; 129(3): 535-7.
  • Barber RD, Harmer DW, Coleman RA, Clark BJ. GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissue. Physiol Genomics 2005; 21(3): 389-95.
  • Becker C, Hammerle-Fickinger A, Riedmaier I, Pfaffl MW. mRNA and microRNA quality control for RT-qPCR analysis. Methods 2010; 50(4):237-43.
  • Blomberg J, Andersson M, Faldt R. Differential pattern of oncogene and beta-actin expression in leukaemic cells from AML patients. J Haematol 1987; 65(1): 83-6.
  • Bogaert L, Van Poucke M, De Baere C, Peelman L, Gasthuys F, Martens A. Selection of a set of reliable reference genes for quantitative real-time PCR in normal equine skin and in equine sarcoids. BMC Biotechnol 2006; 6(24): 1-7.
  • Burns MJ, Nixon GJ, Foy CA, Harris N. Standardisation of data from real-time quantitative PCR methods – evaluation of outliers and comparison of calibration curves. BMC Biotechnol 2005; 5(31): 1-13.
  • Bustin SA, Beaulieu JF, Huggett J, Jaggi R, Kibenge FSB, Olsvik PA, Penning LC, Toegel S. MIQE precis: Practical implementation of minimum standard quidelines for fluorescence based quantitative real-time PCR expreriments. BMC Mol Biol 2010; 11(74): 1-5.
  • Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R,T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT. The MIQE Guidelines: Minimum information for publication of quantitive real-time PCR experiments. Clin Chem 2009; 55(4): 611-22.
  • Bustin SA. Why the need for qPCR publication guidelines?-The case for MIQE. Methods 2010b; 50(4): 217-26.
  • Chervoneva I, Li Y, Schulz S, Croker S, Wilson C, Waldman SA, Hyslop T. Selection of optimal reference genes for normalization in quantitative RT-PCR. BMC Bioinforma 2010; 11(253): 1-15.
  • Cinar MU, Islam MA, Uddin MJ, Tholen E, Tesfaye D, Looft C, Schellander K. Evaluation of suitable reference genes for gene expression studies in porcine alveolar macrophages in response to LPS and LTA. BMC Res Notes 2012; 5(107): 1-14.
  • Cross NC, Lin F, Goldman JM. Appropriate controls for reverse transcription polymerase chain reaction (RT-PCR). Br J Haematol 1994; 87(1): 218.
  • De Spiengelaere W, Dern-Wieloch J, Weigel R, Schumacher V, Schorle H, Nettersheim D, Bergmann M, Brehm R, Kliesch S, Vandekerckhove L, Fink C. Reference gene validation for RT-PCR, a note on different available software packages. PLoS One 2015; 10(3): e122515.
  • Du M, Wang YW, Yue PY, Zhou PY, Yao W, Li X, Ding XB, Liu XF, Guo H, Ma WZ. Selection of reference genes in canine uterine tissues. Genet Mol Res 2016; 15(2): 1-14. Eisenberg E, Levanon EY. Human housekeeping genes are compact. Trends Genet 2003; 19(7): 362-5.
  • Farre D, Bellora N, Mularoni L, Messeguer X, Alba MM. Housekeeping genes tend to Show reduced upstream sequence conservation. Genome Bio 2007; 8(7): RI40. Fleige S, Pfaffl MW. RNA integrity and the effect on the real-time qRT-PCR performance. Mol Aspects Med 2006; 27(2-3): 126-39.
  • Goldman M. Housekeeping Gene. Brenner S. Miller JH. eds. In: Encyclopedia of Genetics. Academic Press. 2001; pp. 978.
  • Gong S, Schmotzer CL, Zhou L. Evaluation of quantitative real-time PCR as a hepatitis C virus supplementary test after RIBA discontinuation. J Clin Lab Anal 2016; 30(5): 418-23.
  • Hashemipetroudi SH, Nematzadeh G, Ahmadian G, Yamchi A, Kuhlmann M. Assesment of DNA contamination in RNA samples based on ribosomal DNA. J Vis Exp 2018; 131(22): e55451.
  • Hayashi R. Gene expression and the impact of antioxidant supplements in the cataractous lens. Preedy VR. eds. Handbook of Nutrition, Diet and the Eye. London: Academic press 2014; p. 517-24.
  • Huggett J, Dheda K, Bustin S, Zumla A. Real-Time RT-PCR normalisation; strategies and considerations. Genes Immun 2005; 6(4): 279-84.
  • Jarczak J, Kaba J, Bagnicka E. The validation of housekeeping genes as a reference in quantitative Real Time PCR analysis: application in the milk somatic cells and frozen whole blood of goats infected with caprine arthritis encephalitis virüs. Gene 2014; 549(2): 280-5.
  • Kaur R, Sodhi M, Sharma A, Sharmi VL, Verma P, Swami SK, Kumari P, Mukesh M. Selection of suitable reference genes for normalizastion of quantitative RT-PCR expression data across twelve tissues of riverine buffaloes. PLoS One 2018; 13(3): E01191668.
  • Klatte M, Bauer P. Accurate Real-time reverse transcription quantitative PCR. Methods Mol Biol 2009; 479(4): 61-77.
  • Korkmaz Agaoglu O, Agaoglu AR, Ozmen O, Saatcı M, Schäfer-Somi S, Aslan S. Investigation of Insulin-like Growth Factor Gene Family in Cat Uterus During Pregnancy at mRNA and Protein Level. XXth International Congress of the European Veterinary Society for Small Animal Reproduction (EVSSAR), June 29-July 1, 2017; Vienna-Austria.
  • Kozera B, Rapacz M. Reference genes in real-time PCR. J Appl Genetics 2013; 54(4): 391-406.
  • Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitive PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25(4): 402-8.
  • Mahoney DJ, Carey K, Fu MH, Snow R, Cameron-Smith D, Parise G, Tarnopolsky MA. Real-time RT-PCR analysis of housekeeping genes in human skeletal muscle following acute exercise. 2004; 18(2): 226-31.
  • Nolan T, Bustin S. Procedures for Quality Control of RNA Samples for Use in Quantitative Reverse Transcription PCR. Keen JT Birch L. eds. In: Essentials of nucleic acid analysis: a robust approach. London: The royal society of Chemistry, 2008; pp. 189-207.
  • Panina Y, Germond A, Masui S, Watanabe TM. Validation of common housekeeping genes as reference for qPCR gene expression analysis during IPS reprogramming process. Sci Rep 2018; 8(1): 8716.
  • Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 2002; 30(9): e36.
  • Pfaffl MW, Tichopad A, Prgomet C, Neuvians TP. Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper-Excel-based tool using pair-wise correlations. Biotech Lett 2004; 26(6): 509-15.
  • Pfaffl MW. A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res 2001; 29(9): e45.
  • Sadek KH, Cagampang FR, Bruce KD, Shreeve N, Macklon N, Cheong Y. Variation in stability of housekeeping genes in endometrium of healthy and polycystic ovarian syndrome women. Hum Reprod 2012; 27(1): 251-6.
  • Segundo-Val IS, Sanz-Lozano CS. Introduction to the gene expression analysis. Methods Mol Biol 2016; 1434(3): 29-43.
  • Sikand K, Singh J, Ebron JS, Shukla GC. Housekeeping gene selection advisory: Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) and β-Actin are targets of miR-644a. PLoS One 2012; 7(10): e47510.
  • Ståhlberg A, Hakansson J, Xian X, Semb H, Kubista M. Properties of the reverse transcription reaction in mRNA quantification. Clin Chem 2004; 50(3): 509-15.
  • Sullivan-Gunn M, Hinch E, Vaughan V, Lewandowski P. Choosing a stable housekeeping gene and protein is essential in generating valid gene and protein expression results. Br J Cancer 2011; 104(6): 1055.
  • Tilli TM, Castro CS, Tuszynski JA, Carels N. A strategy to identify housekeeping genes suitable for analysis in breast cancer diseases. BMC Genomics 2016; 17(1): 639.
  • Uddin M, Cinar M, Tesfaye D, Looft C, Tholen E, Schellander K. Age-related changes in relative expression stability of commonly used housekeeping genes in selected porcine tissues. BMC Res Notes. 2011; 4(441): 1-13.
  • Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002; 3(7): 1-12.
  • VanGuilder HD, Vrana KE, Freeman WM. Twenty-five years of quantitative PCR for gene expression analysis. Biotechniques 2008; 44(5): 619-26.
  • Xie F, Sun G, Stiller JW and Zhang B. Genome-wide functional analysis of the cotton transcriptome by creating an integrated EST database. PLoS One 2011; 6(11): e26980.
  • Zhong H, Simons JW. Direct comparison of GAPDH, beta-actin, cyclophilin, and 28S rRNA as internal standards for quantifying RNA levels under hypoxia. Biochem Biophys Res Commun 1999; 259(3): 523-6.
Toplam 45 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Bölüm Derlemeler
Yazarlar

Özgecan Korkmaz Ağaoğlu 0000-0002-7414-1725

Özge Sidekli Bu kişi benim

Yayımlanma Tarihi 23 Mart 2020
Gönderilme Tarihi 15 Nisan 2019
Kabul Tarihi 23 Temmuz 2019
Yayımlandığı Sayı Yıl 2020 Cilt: 17 Sayı: 1

Kaynak Göster

APA Korkmaz Ağaoğlu, Ö., & Sidekli, Ö. (2020). Kantitatif RT-PCR (RT-qPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi ve Validasyonu. Erciyes Üniversitesi Veteriner Fakültesi Dergisi, 17(1), 76-83. https://doi.org/10.32707/ercivet.655015
AMA Korkmaz Ağaoğlu Ö, Sidekli Ö. Kantitatif RT-PCR (RT-qPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi ve Validasyonu. Erciyes Üniv Vet Fak Derg. Mart 2020;17(1):76-83. doi:10.32707/ercivet.655015
Chicago Korkmaz Ağaoğlu, Özgecan, ve Özge Sidekli. “Kantitatif RT-PCR (RT-QPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi Ve Validasyonu”. Erciyes Üniversitesi Veteriner Fakültesi Dergisi 17, sy. 1 (Mart 2020): 76-83. https://doi.org/10.32707/ercivet.655015.
EndNote Korkmaz Ağaoğlu Ö, Sidekli Ö (01 Mart 2020) Kantitatif RT-PCR (RT-qPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi ve Validasyonu. Erciyes Üniversitesi Veteriner Fakültesi Dergisi 17 1 76–83.
IEEE Ö. Korkmaz Ağaoğlu ve Ö. Sidekli, “Kantitatif RT-PCR (RT-qPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi ve Validasyonu”, Erciyes Üniv Vet Fak Derg, c. 17, sy. 1, ss. 76–83, 2020, doi: 10.32707/ercivet.655015.
ISNAD Korkmaz Ağaoğlu, Özgecan - Sidekli, Özge. “Kantitatif RT-PCR (RT-QPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi Ve Validasyonu”. Erciyes Üniversitesi Veteriner Fakültesi Dergisi 17/1 (Mart 2020), 76-83. https://doi.org/10.32707/ercivet.655015.
JAMA Korkmaz Ağaoğlu Ö, Sidekli Ö. Kantitatif RT-PCR (RT-qPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi ve Validasyonu. Erciyes Üniv Vet Fak Derg. 2020;17:76–83.
MLA Korkmaz Ağaoğlu, Özgecan ve Özge Sidekli. “Kantitatif RT-PCR (RT-QPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi Ve Validasyonu”. Erciyes Üniversitesi Veteriner Fakültesi Dergisi, c. 17, sy. 1, 2020, ss. 76-83, doi:10.32707/ercivet.655015.
Vancouver Korkmaz Ağaoğlu Ö, Sidekli Ö. Kantitatif RT-PCR (RT-qPCR) Çalışmalarında Uygun Housekeeping Genlerin (HKGs) Seçimi ve Validasyonu. Erciyes Üniv Vet Fak Derg. 2020;17(1):76-83.

Cited By

Commonly Used Normalization Approaches in MicroRNA Expression Profiling
Erciyes Üniversitesi Veteriner Fakültesi Dergisi
https://doi.org/10.32707/ercivet.1142293
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https://dergipark.org.tr/tr/download/journal-file/20610

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