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Hereditas(Beijing) ›› 2018, Vol. 40 ›› Issue (3): 227-236.doi: 10.16288/j.yczz.17-369

• Reviews • Previous Articles     Next Articles

Co-amplification at lower denaturation temperature-PCR: methodology and applications

Hui Liang1,2,Guojie Chen3,Yan Yu4,Likuan Xiong1,2()   

  1. 1. Central Laboratory, Bao’an Maternal and Child Health Hospital, Jinan University, Shenzhen 518102, China;
    2. Shenzhen Key Laboratory of Birth Defects, Shenzhen 518102, China
    3. Department of Gastroenterology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
    4. Department of Obstetrics, Bao’an Maternal and Child Health Hospital, Jinan University, Shenzhen 518102, China;
  • Received:2017-11-06 Revised:2018-02-02 Online:2018-03-20 Published:2018-02-08
  • Supported by:
    [Supported by Sanming Project of Medicine in Shenzhen-Brith Defects Prevention Research and Transformation Team (No. SZSM201406007), Shenzhen Key Laboratory of Birth Defects (No. ZDSYS201504301707152 and Science and Technology Plan Project of Baoan District (Nos. 2014067, 2017JD001)]

Abstract:

Co-amplification at lower denaturation temperature-polymerase chain reaction (COLD-PCR) is a novel form of PCR that selectively denatures and amplifies low-abundance mutations from mixtures of wild-type and mutation-containing sequences, enriching the mutation 10 to 100 folds. Due to the slightly altered melting temperature (Tm) of the double-stranded DNA and the formation of the mutation/wild-type heteroduplex DNA, COLD-PCR methods are sensitive, specific, accurate, cost-effective and easy to maneuver, and can enrich mutations of any type and at any position, even unknown mutations within amplicons. COLD-PCR and its improved methods are now applied in cancer, microorganisms, prenatal screening, animals and plants. They are extremely useful for early diagnosis, monitoring the prognosis of disease and the efficiency of the treatment, drug selection, prediction of prognosis, plant breeding and etc. In this review, we introduce the principles, key techniques, derived methods and applications of COLD-PCR.

Key words: co-amplification at lower denaturation temperature polymerase chain reaction (COLD-PCR), Tc, selectively amplifies, low-abundance mutation, detection threshold