The Polymerase Chain Reaction (PCR) is a well-known approach to amplify a specific DNA sequence. PCR involves the reiterative cycling of a reaction cocktail between different temperatures to achieve amplification. As routine as PCR is in the molecular biology and molecular diagnostic laboratory, there are other methods of sequence-specific DNA amplification. These alternative approaches often do not require changing the reaction temperature and are, therefore, often referred to as sequence-specific isothermal amplification protocols. Isothermal amplification protocols are varied and have varied advantages. However, some common advantages are that isothermal techniques are extremely fast and they do not require thermocyclers.
Four examples of sequence-specific isothermal DNA amplification technologies include:
- Loop-mediated Isothermal Amplification (LAMP)
- 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)
- Typical cDNA Synthesis Protocol
- Typical LAMP Protocol (M0275)
- Typical LAMP Protocol (M0374)
- Typical LAMP Protocol (M0537)
- Typical LAMP Protocol (M0538)
- Typical RT-LAMP Protocol
- WarmStart Colorimetric LAMP 2X Master Mix Typical LAMP Protocol (M1800)
- WarmStart LAMP Kit (DNA & RNA) Protocol (E1700)
Anatomy of a Polymerase - How Structure Effects Function
Understanding Variability in DNA Amplification Reactions
- Manabu Nemoto, Yoshinori Morita, Hidekazu Niwa, Hiroshi Bannai, Koji Tsujimura, Takashi Yamanaka, Takashi Kondo 2015. Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification. J Virol Methods. 215-216, PubMedID: 25682750, DOI: 10.1016/j.jviromet.2015.02.001
- Sanchita Bhadra, Yu Sherry Jiang, Mia R Kumar, Reed F Johnson, Lisa E Hensley, Andrew D Ellington 2015. Real-Time Sequence-Validated Loop-Mediated Isothermal Amplification Assays for Detection of Middle East Respiratory Syndrome Coronavirus (MERS-CoV). PLoS One. 10, PubMedID: 25856093, DOI: 10.1371/journal.pone.0123126
- Trisadee Khamlor, Petai Pongpiachan, Rangsun Parnpai, Kanchana Punyawai, Siwat Sangsritavong, Nipa Chokesajjawatee 2015. Bovine embryo sex determination by multiplex loop-mediated isothermal amplification. Theriogenology. 83, PubMedID: 25542460, DOI: 10.1016/j.theriogenology.2014.11.025
- DoKyung Lee, Eun Jin Kim, Paul E Kilgore, Soon Ae Kim, Hideyuki Takahashi, Makoto Ohnishi, Dang Duc Anh, Bai Qing Dong, Jung Soo Kim, Jun Tomono, Shigehiko Miyamoto, Tsugunori Notomi, Dong Wook Kim, Mitsuko Seki 2015. Clinical Evaluation of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Neisseria meningitidis in Cerebrospinal Fluid. PLoS One. 10, PubMedID: 25853422, DOI: 10.1371/journal.pone.0122922
- Mohammad Reza Allahyar Torkaman, Kazunari Kamachi, Vajihe Sadat Nikbin, Masoumeh Nakhost Lotfi, Fereshteh Shahcheraghi 2015. Comparison of loop-mediated isothermal amplification and real-time PCR for detecting Bordetella pertussis. J Med Microbiol. 64, PubMedID: 25596118, DOI: 10.1099/jmm.0.000021
- Aongart Mahittikorn, Hirotake Mori, Supaluk Popruk, Amonrattana Roobthaisong, Chantira Sutthikornchai, Khuanchai Koompapong, Sukhontha Siri, Yaowalark Sukthana, Duangporn Nacapunchai 2015. Development of a Rapid, Simple Method for Detecting Naegleria fowleri Visually in Water Samples by Loop-Mediated Isothermal Amplification (LAMP). PLoS One. 10, PubMedID: 25822175, DOI: 10.1371/journal.pone.0120997
- Tanner NA, Evans TC Jr. 2014. Loop-mediated isothermal amplification for detection of nucleic acids Curr Protoc Mol Biol. 105, PubMedID: 24510439, DOI:
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