scholarly journals Importance of housekeeping gene selection for accurate reverse transcription-quantitative polymerase chain reaction in a wound healing model

2010 ◽  
Vol 18 (5) ◽  
pp. 460-466 ◽  
Author(s):  
Anna Turabelidze ◽  
Shujuan Guo ◽  
Luisa A. DiPietro
Keyword(s):  
Polymerase Chain Reaction ◽  
Wound Healing ◽  
Gene Selection ◽  
Quantitative Polymerase Chain Reaction ◽  
Housekeeping Gene ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Wound Healing Model ◽  
Selection For ◽  
Healing Model

Reference gene selection for quantitative real-time polymerase chain reaction in Populus

2011 ◽  
Vol 408 (2) ◽  
pp. 337-339 ◽  
Author(s):  
Meng Xu ◽  
Bo Zhang ◽  
Xiaohua Su ◽  
Shougong Zhang ◽  
Minren Huang
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Reference Gene ◽  
Gene Selection ◽  
Chain Reaction ◽  
Reference Gene Selection ◽  
Polymerase Chain ◽  
Selection For

scholarly journals Quick and Accurate Detection and Quantification of Magnaporthe oryzae in Rice Using Real-Time Quantitative Polymerase Chain Reaction

Plant Disease ◽  
2015 ◽  
Vol 99 (2) ◽  
pp. 219-224 ◽  
Author(s):  
Geng Sun ◽  
Jinliang Liu ◽  
Guihua Li ◽  
Xianghui Zhang ◽  
Tingting Chen ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Magnaporthe Oryzae ◽  
Quantitative Polymerase Chain Reaction ◽  
Housekeeping Gene ◽  
Practical Significance ◽  
Rice Seed ◽  
Chain Reaction ◽  
Pure Cultures ◽  
Polymerase Chain

Rice blast, caused by Magnaporthe oryzae, is one of the most severe fungal diseases in rice worldwide. In this study, we developed methods to quickly and accurately detect and quantify M. oryzae in the pure cultures of the fungus, rice plants, and rice seed by using SYBR Green I of the real-time quantitative polymerase chain reaction (qPCR). Results of absolute qPCR show that Magnaporthe oryzae can be detected at as low as 6.9 × 10−5 ng of genomic DNA. Results also show that all 10 varieties of rice seed examined in this study contain this fungus, indicating that M. oryzae is generally widespread in rice seed. We report the quantification of DNA of M. oryzae in rice leaves collected in the field, instead of in the lab, using relative qPCR by using rice actin gene as a housekeeping gene. Our results show great practical significance because we would know the potential fungal infection even before planting.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

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