Literature DB >> 26514670

A powerless on-the-spot detection protocol for transgenic crops within 30 min, from leaf sampling up to results.

Liu Wang1, Rui Wang1, Yonghua Yu1, Fang Zhang1, Xiaofu Wang2, Yibin Ying1, Jian Wu3, Junfeng Xu2.   

Abstract

The requirement of power-dependent instruments or excessive operation time usually restricts current nucleic acid amplification methods from being used for detection of transgenic crops in the field. In this paper, an easy and rapid detection method which requires no electricity supply has been developed. The time-consuming process of nucleic acid purification is omitted in this method. DNA solution obtained from leaves with 0.5 M sodium hydroxide (NaOH) can be used for loop-mediated isothermal amplification (LAMP) only after simple dilution. Traditional instruments like a polymerase chain reaction (PCR) amplifier and water bath used for DNA amplification are abandoned. Three kinds of dewar flasks were tested and it turned out that the common dewar flask was the best. Combined with visual detection of LAMP amplicons by phosphate (Pi)-induced coloration reaction, the whole process of detection of transgenic crops via genetically pure material (leaf material of one plant) could be accomplished within 30 min. The feasibility of this method was also verified by analysis of practical samples.

Entities:  

Keywords:  Crude DNA extract; Loop-mediated isothermal amplification; Transgenic crop detection; Unpowered device; Visual detection

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Year:  2015        PMID: 26514670     DOI: 10.1007/s00216-015-9128-x

Source DB:  PubMed          Journal:  Anal Bioanal Chem        ISSN: 1618-2642            Impact factor:   4.142


  2 in total

1.  Rapid sex identification method of spinach (Spinacia oleracea L.) in the vegetative stage using loop-mediated isothermal amplification.

Authors:  Naoko Fujita; Yu Ayukawa; Mitsutoshi Fuke; Tohru Teraoka; Kyoko Watanabe; Tsutomu Arie; Ken Komatsu
Journal:  Planta       Date:  2016-11-12       Impact factor: 4.116

2.  Rapid high throughput template preparation (rHTTP) method: a novel cost effective method of direct PCR for a wide range of plants.

Authors:  Prassan Choudhary; Sudipta Das; Hillol Chakdar; Arjun Singh; Sanjay Kumar Goswami; Anil Kumar Saxena
Journal:  BMC Biotechnol       Date:  2019-10-26       Impact factor: 2.563

  2 in total

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