Shortening distance of forward and reverse primers for nucleic acid isothermal amplification
-
Qu Haitao
,
Zhang Wenchao
Abstract
Existent nucleic acid isothermal detection techniques for clinical diseases are difficult to promote greatly due to limitations in such aspects as methodology, costs of detection, amplification efficiency and conditions for operation. There is therefore an urgent need for a new isothermal amplification method with the characteristics of high accuracy, easy operation, short time of detection and low costs. We have devised a new method of nucleic acid isothermal amplification using Bst DNA polymerase under isothermal conditions (60–65°C). We call this method of amplification by shortening the distance between forward and reverse primers for nucleic acid isothermal amplification SDAMP. The results demonstrated that this technique is highly sensitive, specific and has short reaction times (40–60 min). Results of sequencing show that the products of SDAMP amplification are mainly polymers formed by series connection of monomers formed through linkage of forward primer and complementary sequences in reverse primer via a few bases. The method is different from current methods of nucleic acid amplification. Our study shows, however, that it is a specific method of nucleic acid isothermal amplification depending on interactions between primers and DNA template.
Acknowledgments
This work was supported by Heilongjiang Prevention and Treatment of Infectious Diseases Hospital and Huazhong Agricultural University.
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Artikel in diesem Heft
- Frontmatter
- Reviews
- The top skin-associated genes: a comparative analysis of human and mouse skin transcriptomes
- Human dyskerin: beyond telomeres
- Assembly and function of small RNA – Argonaute protein complexes
- Minireview
- The globin superfamily: functions in nitric oxide formation and decay
- Research Articles/Short Communications
- Protein Structure and Function
- N-homocysteinylation of apolipoprotein A-I impairs the protein’s antioxidant ability but not its cholesterol efflux capacity
- Lucimycin, an antifungal peptide from the therapeutic maggot of the common green bottle fly Lucilia sericata
- A lysine-methionine exchange in a coronavirus surface protein transforms a retention motif into an endocytosis signal
- Cell Biology and Signaling
- Upregulation of the thioredoxin-dependent redox system during differentiation of 3T3-L1 cells to adipocytes
- Novel Techniques
- Shortening distance of forward and reverse primers for nucleic acid isothermal amplification
