Literature DB >> 25697660

Rapid and simple method of qPCR primer design.

Brenda Thornton1, Chhandak Basu.   

Abstract

Quantitative real-time polymerase chain reaction (qPCR) is a powerful tool for analysis and quantification of gene expression. It is advantageous compared to traditional gel-based method of PCR, as gene expression can be visualized "real-time" using a computer. In qPCR, a reporter dye system is used which intercalates with DNA's region of interest and detects DNA amplification. Some of the popular reporter systems used in qPCR are the following: Molecular Beacon(®), SYBR Green(®), and Taqman(®). However, success of qPCR depends on the optimal primers used. Some of the considerations for primer design are the following: GC content, primer self-dimer, or secondary structure formation. Freely available software could be used for ideal qPCR primer design. Here we have shown how to use some freely available web-based software programs (such as Primerquest(®), Unafold(®), and Beacon designer(®)) to design qPCR primers.

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Year:  2015        PMID: 25697660     DOI: 10.1007/978-1-4939-2365-6_13

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  16 in total

1.  Molecular Mechanism and Genetic Determinants of Buprofezin Degradation.

Authors:  Xueting Chen; Junbin Ji; Leizhen Zhao; Jiguo Qiu; Chen Dai; Weiwu Wang; Jian He; Jiandong Jiang; Qing Hong; Xin Yan
Journal:  Appl Environ Microbiol       Date:  2017-08-31       Impact factor: 4.792

2.  Suggested guidelines for validation of real-time PCR assays in veterinary diagnostic laboratories.

Authors:  Kathy Toohey-Kurth; Monica M Reising; Rebecca L Tallmadge; Laura B Goodman; Jianfa Bai; Steven R Bolin; Janice C Pedersen; Mangkey A Bounpheng; Roman M Pogranichniy; Jane Christopher-Hennings; Mary Lea Killian; Donna M Mulrooney; Roger Maes; Shri Singh; Beate M Crossley
Journal:  J Vet Diagn Invest       Date:  2020-09-28       Impact factor: 1.279

3.  Experimental Design for Time-Series RNA-Seq Analysis of Gene Expression and Alternative Splicing.

Authors:  Nikoleta A Tzioutziou; Allan B James; Wenbin Guo; Cristiane P G Calixto; Runxuan Zhang; Hugh G Nimmo; John W S Brown
Journal:  Methods Mol Biol       Date:  2022

4.  Mechanisms of Sporicidal Activity Induced by Ionized Hydrogen Peroxide in the Spores of Bacillus atrophaeus.

Authors:  Miguel A Grimaldo; Nicole L Mendell; Nathen E Bopp; Donald H Bouyer; Claudio Mafra
Journal:  Appl Biosaf       Date:  2021-09-13

5.  Human Trabecular Meshwork (HTM) Cells Treated with TGF-β2 or Dexamethasone Respond to Compression Stress in Different Manners.

Authors:  Megumi Watanabe; Tatsuya Sato; Yuri Tsugeno; Araya Umetsu; Soma Suzuki; Masato Furuhashi; Yosuke Ida; Fumihito Hikage; Hiroshi Ohguro
Journal:  Biomedicines       Date:  2022-06-06

6.  Ectopic Overexpression of PPARγ2 in the Heart Determines Differences in Hypertrophic Cardiomyopathy After Treatment With Different Thiazolidinediones in a Mouse Model of Diabetes.

Authors:  Xuemei Cao; Min Mao; Junlin Diao; Yi Hou; Hong Su; Yongjun Gan; Jibin Li; Xiaoyong Tong; Chaodong Wu; Zhong Zuo; Xiaoqiu Xiao
Journal:  Front Pharmacol       Date:  2021-07-07       Impact factor: 5.810

7.  Normalization for Relative Quantification of mRNA and microRNA in Soybean Exposed to Various Abiotic Stresses.

Authors:  Weican Liu; Yu Deng; Yonggang Zhou; Huan Chen; Yuanyuan Dong; Nan Wang; Xiaowei Li; Aysha Jameel; He Yang; Min Zhang; Kai Chen; Fawei Wang; Haiyan Li
Journal:  PLoS One       Date:  2016-05-13       Impact factor: 3.240

8.  Selection of Valid Reference Genes for Reverse Transcription Quantitative PCR Analysis in Heliconius numata (Lepidoptera: Nymphalidae).

Authors:  Florence Piron Prunier; Mathieu Chouteau; Annabel Whibley; Mathieu Joron; Violaine Llaurens
Journal:  J Insect Sci       Date:  2016-06-07       Impact factor: 1.857

9.  RgC3H Involves in the Biosynthesis of Allelopathic Phenolic Acids and Alters Their Release Amount in Rehmannia glutinosa Roots.

Authors:  Yanhui Yang; Zhongyi Zhang; Ruifang Li; Yanjie Yi; Heng Yang; Chaojie Wang; Zushiqi Wang; Yunyi Liu
Journal:  Plants (Basel)       Date:  2020-04-29

10.  Primer set 2.0 for highly parallel qPCR array targeting antibiotic resistance genes and mobile genetic elements.

Authors:  Robert D Stedtfeld; Xueping Guo; Tiffany M Stedtfeld; Hongjie Sheng; Maggie R Williams; Kristin Hauschild; Santosh Gunturu; Leo Tift; Fang Wang; Adina Howe; Benli Chai; Daqiang Yin; James R Cole; James M Tiedje; Syed A Hashsham
Journal:  FEMS Microbiol Ecol       Date:  2018-09-01       Impact factor: 4.194

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