Detection and image processing after nucleic acid amplification in emulsion

Tupik, A. N.; Novikov, Lev V.; Evstrapov, A. A.

doi:10.1088/1742-6596/1400/3/033009

Cited by 1 publication

(1 citation statement)

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“…Reaction compartments can be very simply generated as water in oil emulsions or by using an array of sealed chambers. The compartments are seeded in a limited dilution step ensuring that less than one template is present in each compartment (Tupik et al 2019). Thus, the rare occurrence of a rapidly amplifying artifact or contamination is contained in one compartment and cannot supersede the desired amplicons in other compartments.…”

Section: Remedies For Artifacts and Contaminationsmentioning

confidence: 99%

Isothermal amplifications – a comprehensive review on current methods

Glökler

Lim

Ida

et al. 2021

Critical Reviews in Biochemistry and Molecular Biology

102

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The introduction of nucleic acid amplification techniques has revolutionized the field of medical diagnostics in the last decade. The advent of PCR catalyzed the increasing application of DNA, not just for molecular cloning but also for molecular based diagnostics. Since the introduction of PCR, a deeper understanding of molecular mechanisms and enzymes involved in DNA/RNA replication has spurred the development of novel methods devoid of temperature cycling. Isothermal amplification methods have since been introduced utilizing different mechanisms, enzymes, and conditions. The ease with which isothermal amplification methods have allowed nucleic acid amplification to be carried out has had a profound impact on the way molecular diagnostics are being designed after the turn of the millennium. With all the advantages isothermal amplification brings, the issues or complications surrounding each method are heterogeneous making it difficult to identify the best approach for an end-user. This review pays special attention to the various isothermal amplification methods by classifying them based on the mechanistic characteristics which include reaction formats, amplification information, promoter, strand break, and refolding mechanisms. We would also compare the efficiencies and usefulness of each method while highlighting the potential applications and detection methods involved. This review will serve as an overall outlook on the journey and development of isothermal amplification methods as a whole.

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Section: Remedies For Artifacts and Contaminationsmentioning

confidence: 99%