Isothermal Amplification & Strand Displacement
Loop-mediated isothermal amplification (LAMP) uses 4-6 primers recognizing 6-8 distinct regions of target DNA. A strand-displacing DNA polymerase initiates synthesis and 2 of the primers form loop structures to facilitate subsequent rounds of amplification.
Strand Displacement Amplification (SDA) utilizes two outer “bump” primers and two inner primers with 5’ tail regions that contain a nicking enzyme recognition site. In conjunction with a nicking enzyme (e.g., Nt.BstNBI), amplification of discrete DNA products occurs in rapid fashion.
Choose Type:
- One-Step qHDA (Real-time quantitative tHDA)
- One-Step qRT-HDA (Real-time quantitative RT-HDA)
- One-Step RT-HDA (Reverse Transcription tHDA)
- One-Step tHDA (thermostable HDA)
- Two-Step qHDA (Real-time quantitative tHDA)
- Two-Step qRT-HDA (Real-time quantitative RT-HDA)
- Two-Step RT-HDA (Reverse Transcription tHDA)
- Two-Step tHDA (thermostable HDA)
- A-Tailing with Klenow Fragment (3'→5' exo-)
- Typical LAMP Protocol (M0275)
- Typical LAMP Protocol (M0538)
- Typical LAMP Protocol (M0374)
- WarmStart Colorimetric LAMP 2X Master Mix Typical LAMP Protocol (M1800)
- WarmStart LAMP Kit (DNA & RNA) Protocol (E1700)
- Bst-XT WarmStart™ Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) (NEB #M1712)
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Anatomy of a Polymerase - How Function and Structure are Related
Read about the relationship between Polymerase structure and function when copying DNA.
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Using aptamers to control enzyme activity: Hot Start Taq and beyond
This article describes the use of engineered oligonucleotides known as aptamers in regulating the enzyme activity of polymerases and reverse transcriptases.
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Many Initiatives Turning to RT-LAMP as Alternative to PCR for Rapid COVID-19 Screening Assays
There is an increasing demand for large-scale screening and surveillance testing, teams are now scaling up rapid SARS-CoV-2 screening assays incorporating RT-LAMP.
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BMG LABTECH: LAMP assay for detecting SARS-CoV-2 RNA using an absorbance-based colorimetric readout
There is an increasing demand for large-scale screening and surveillance testing, teams are now scaling up rapid SARS-CoV-2 screening assays incorporating RT-LAMP.
- Isothermal Amplification Brochure
Feature Articles
Brochures
- Nzelu CO, Kato H, Peters NC. (2019) Loop-mediated isothermal amplification (LAMP): An advanced molecular point-of-care technique for the detection of Leishmania infection PLoS Negl Trop Dis; 13(11):e0007698, PubMedID: 31697673, DOI: 10.1371/journal.pntd.0007698
- Poole, C.B., Sinha, A., Ettwiller, L., Apone, L., McKay, K., Panchapakesa, V., Lima, N.F., Ferreira, M.U., Wanji, S., Carlow, C.K.S (2019) In silico identification of novel biomarkers and development of new rapid diagnostic tests for the filarial parasites Mansonella perstans and Mansonella ozzardi Scientific Reports; 9-1, 10275. PubMedID: 31311985, DOI: 10.1038/s41598-019-46550-9
- Toldrà A, O'Sullivan CK, Campàs M. (2019) Detecting Harmful Algal Blooms with Isothermal Molecular Strategies Trends Biotechnol; 37(12):1278–1281, PubMedID: 31399265, DOI: 10.1016/j.tibtech.2019.07.003
- Zhang M, Ye J, He JS, et al. (2020) Visual detection for nucleic acid-based techniques as potential on-site detection methods. A review. Anal Chim Acta; 1099:1–15, PubMedID: 31986265, DOI: 10.1016/j.aca.2019.11.056
- Tanner NA, Evans TC Jr. (2014) Loop-mediated isothermal amplification for detection of nucleic acids Curr Protoc Mol Biol; 105, PubMedID: 24510439
- Calvert AE, Biggerstaff BJ, Tanner NA, Lauterbach M, Lanciotti RS. (2017) Rapid colorimetric detection of Zika virus from serum and urine specimens by reverse transcription loop-mediated isothermal amplification PLoS One; 12(9):e0185340, PubMedID: 28945787, DOI: 10.1371/journal.pone.0185340
Template/product specificity: Is RNA or DNA involved? Is the 3´ terminus at a gap, nick or at the end of the template?
Removal of existing nucleotides: Will the nucleotide(s) be removed from the existing polynucleotide chain as part of the protocol? If so, will they be removed from the 5´ or the 3´ end?
Thermal stability: Does the polymerase need to survive incubation at high temperature or is heat inactivation desirable?
Fidelity: Will subsequent sequence analysis or expression depend on the fidelity of the synthesized products?
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