Rapid identification of Takifugu genus using visual loop‐mediated isothermal amplification

Author(s):  
Ruibin Xie ◽  
Hui Li ◽  
Wenjie Yu ◽  
Nan Wang ◽  
Juan Zhang ◽  
...  
2011 ◽  
Vol 47 (No. 4) ◽  
pp. 140-148 ◽  
Author(s):  
N. Rostamkhani ◽  
A. Haghnazari ◽  
M. Tohidfar ◽  
A. Moradi

In an attempt to speed up the process of screening of transgenic cotton (G. hirsutum L.) plants, a visual and rapid loop-mediated isothermal amplification (LAMP) assay was adopted. Genomic DNA was extracted from fresh leaf tissues of T<sub>2</sub> transgenic cotton containing chitinase (chi) and cry1A(b) genes. Detection of genes of interest was performed by polymerase chain reaction (PCR), LAMP and real-time PCR methods. In LAMP assay the amplification was performed after 30 min at 65&deg;C when loop primers were involved in the reaction. The involvement of loop primers decreased the time needed for amplification. By testing serial tenfold dilutions (10<sup>&ndash;1</sup> to 10<sup>&ndash;8</sup>) of the genes of interest, the detection sensitivity of LAMP was found to be 100-fold higher than that of PCR. The rapid DNA extraction method and LAMP assay can be performed within 30 min and the derived LAMP products can be directly observed as visually detectable based on turbidity in the reaction tube. The accuracy of LAMP method in the screening of transgenes was confirmed by PCR and real-time PCR. The developed method was efficient, rapid and sensitive in the screening of cotton transgenic plants. This method can be applied to any other crops.


2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Masashi Kitamura ◽  
Akira Kazato ◽  
Tadashi Yamamuro ◽  
Hirokazu Ando ◽  
Yohei Sasaki ◽  
...  

Crop Science ◽  
2020 ◽  
Vol 60 (6) ◽  
pp. 3004-3012
Author(s):  
Nara Cristina Teixeira ◽  
Adriane Wendland ◽  
Maythsulene Inácio de Souza Oliveira ◽  
Livia Teixeira Duarte Brandão ◽  
Thiago Lívio Pessoa Oliveira Souza ◽  
...  

2017 ◽  
Vol 10 (7) ◽  
pp. 2325-2331 ◽  
Author(s):  
Ya Shi ◽  
Yan Feng ◽  
Changping Xu ◽  
Zhouheng Xu ◽  
Dongqing Cheng ◽  
...  

2009 ◽  
Vol 156 (1-2) ◽  
pp. 32-36 ◽  
Author(s):  
Namal Perera ◽  
Hiroka Aonuma ◽  
Aya Yoshimura ◽  
Tokiyasu Teramoto ◽  
Hiroshi Iseki ◽  
...  

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