Hot start PCR is a modified form of conventional polymerase chain reaction (PCR) that reduces the presence of undesired products and primer dimers due to non-specific DNA amplification at room (or colder) temperatures.[1][2] Many variations and modifications of the PCR procedure have been developed in order to achieve higher yields; hot start PCR is one of them.[3] Hot start PCR follows the same principles as the conventional PCR - in that it uses DNA polymerase to synthesise DNA from a single stranded template.[4] However, it utilizes additional heating and separation methods, such as inactivating or inhibiting the binding of Taq polymerase and late addition of Taq polymerase, to increase product yield as well as provide a higher specificity and sensitivity.[5] Non-specific binding and priming or formation of primer dimers are minimized by completing the reaction mix after denaturation.[6] Some ways to complete reaction mixes at high temperatures involve modifications that block DNA polymerase activity in low temperatures,[1][7] use of modified deoxyribonucleotide triphosphates (dNTPs),[8] and the physical addition of one of the essential reagents after denaturation.[9]
Through these additional methods, hot start PCR is able to decrease the amount of non-specific amplifications which naturally occur during lower temperatures – which remains a problem for conventional PCR. These modifications work overall to ensure that specific enzymes in solution will remain inactive or are inhibited until the optimal annealing temperature is reached.[10] Inhibiting formation of non-specific PCR products, especially in early cycles, results in a substantial increase in sensitivity of amplification by PCR. This is of utmost importance in diagnostic applications of PCR or RT-PCR.
^ abSharkey DJ, Scalice ER, Christy KG, Atwood SM, Daiss JL (May 1994). "Antibodies as thermolabile switches: high temperature triggering for the polymerase chain reaction". Bio/Technology. 12 (5): 506–9. doi:10.1038/nbt0594-506. PMID 7764710. S2CID 2885453.
^Paul N, Shum J, Le T (2010). "Hot start PCR". RT-PCR Protocols. Methods in Molecular Biology. Vol. 630. Humana Press. pp. 301–18. doi:10.1007/978-1-60761-629-0_19. ISBN 9781607616283. PMID 20301005.
^Green, Michael R.; Sambrook, Joseph (May 2018). "Hot Start Polymerase Chain Reaction (PCR)". Cold Spring Harbor Protocols. 2018 (5): pdb.prot095125. doi:10.1101/pdb.prot095125. ISSN 1940-3402. PMID 29717052.
^Cite error: The named reference :2 was invoked but never defined (see the help page).
^van Pelt-Verkuil E, van Belkum A, Hays JP, eds. (2008). "Variants and Adaptations of the Standard PCR Protocol". Principles and Technical Aspects of PCR Amplification. Springer Netherlands. pp. 231–276. doi:10.1007/978-1-4020-6241-4_12. ISBN 9781402062414.
^"How is Hot-Start Technology Beneficial For Your PCR". Thermofisher. Retrieved 2019-10-03.
^"DNTP - The School of Biomedical Sciences Wiki". teaching.ncl.ac.uk. Retrieved 2019-10-09.
HotstartPCR is a modified form of conventional polymerase chain reaction (PCR) that reduces the presence of undesired products and primer dimers due...
to most PCR methods are as follows: Initialization: This step is only required for DNA polymerases that require heat activation by hot-startPCR. It consists...
mixture reaches the higher temperatures. These methods are referred to as hot-startPCR. Wax: in this method the enzyme is spatially separated from the reaction...
Le: HotstartPCR. In: Methods Mol Biol. (2010), Volume 630, p. 301–18. PMID 20301005. M. F. Kramer, D. M. Coen: Enzymatic amplification of DNA by PCR: standard...
of amplified product in real time as the amplification progresses. Hot-startPCR is a technique performed manually by heating the reaction components...
prevent non-specific primer interactions, thus enabling successful PCR. A "hot-start" polymerase enzyme whose activity is blocked unless it is heated to...
positions containing a ribonucleotide. A variant of hotstartPCR, known as RNase H-dependent PCR or rhPCR, has been described using a thermostable RNase HII...
with the terms and components used in the PCR process.) The history of the polymerase chain reaction (PCR) has variously been described as a classic...
Ferris, Michael J.; Henson, Joan M. (2003). "PCR Detection and Analysis of the Free-Living Amoeba Naegleria in Hot Springs in Yellowstone and Grand Teton National...
techniques for performing this are whole plasmid single PCR, and overlap extension PCR. Whole plasmid single PCR is also referred to as site directed mutagenesis...
thermal cyclers, used for the synthesis of DNA by polymerase chain reaction (PCR), a common molecular biological technique, which requires the rapid heating...
(which Minaj attended) required proof of vaccination and a negative COVID-19 PCR test to attend. Studio albums Pink Friday (2010) Pink Friday: Roman Reloaded...
entire cards can be replaced while the system is hot, thus minimizing downtime due to hardware failure. PCR consists of a custom OAM interface that is highly...
ancient bacteria could be extracted from the solid inclusions and amplified by PCR. Microbial studies on fluid inclusions are foreseen to attempt to evidence...
Palasek, Stan (23 May 2013). "Primordial RNA Replication and Applications in PCR Technology". arXiv:1305.5581v1 [q-bio.BM]. Vlassov, Alexander V.; Kazakov...
reaction (PCR) tests for Lyme disease have also been developed to detect the genetic material (DNA) of the Lyme disease spirochete. Culture or PCR are the...
can also be confirmed by isolation of viral DNA detected by PCR or direct hybridization. PCR Is considered significantly more sensitive to detecting viral...
assay with a natural hot-start reaction. This created pool of extension products of DNA sequence forms amplicons amplified by PCR where each amplicon sequence...
leprae bacterial DNA can be identified using a polymerase chain reaction (PCR)-based technique. This molecular test alone is not sufficient to diagnose...
diversity of thermophilic cyanobacteria in Rupite hot spring identified by microscopy, Cultivation, Single-cell PCR and Amplicon sequencing". Extremophiles. 23...
which provided training for the leadership of the Romanian Communist Party (PCR) before the 1989 Revolution. The former party secretary from the Ștefan Gheorghiu...
reaction (PCR) process. The retrieval of these bacteria can be achieved with no impact on the ecosystem. Other bacteria in the Yellowstone hot springs may...
In the Romanian general election of 1946, the Romanian Communist Party (PCR) employed widespread intimidation tactics and electoral fraud to obtain 80...
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