eISSN: 2221-6197 DOI: 10.31301/2221-6197

PCR enhancers. I. General information

Year: 2023

Pages: 218-223

Number: Volume 15, issue 3

Type: scientific article

Summary:

PCR is the most popular method for nucleic acids amplification, which characterized by high specificity and sensitivity. PCR is widely used in research and in practice. However, when nucleotide sequences, called “difficult” templates, are amplified, false-negative results can be obtained for samples containing a target in the reaction mixture. The same result can be caused by impurities that are co-extracted with nucleic acids and inhibit DNA polymerase. To eliminate this, substances called PCR enhancers are added to the reaction mixture, which can stabilize the DNA polymerase and/or, on the contrary, destabilize hydrogen bonds in GC-rich regions. The use of proteins, amino acids, carbohydrates, polyhydric alcohols, amides, sulfones, sulfoxides, nonionic detergents, zwitterionic compounds, various nanoparticles, as well as

Keywords:

DNA, RNA, nucleic acids, PCR, PCR enhancers, difficult template, GC-rich sequences, long PCR

References:

1. Akhmetzianova L.U., Davletkulov T.M., Sakhabutdinova A.R., Chemeris A.V., Gubaydullin I.M., Garafutdinov R.R. LAMPrimers iQ: New primer design software for loop-mediated isothermal amplification (LAMP). Anal. Biochem. 2023. V. 684. 115376. doi: 10.1016/j.ab.2023.115376

2. Chemeris A.V., Chemeris D.A., Magdanov E.G., Garafutdinov R.R., Nagaev N.R., Vakhitov V.A. Causes of false-negative PCR and how to avoid some of them. Biomics. 2012. V. 4. P. 31-47. (In Russian)

3. Chemeris A.V., Magdanov E.G., Garafutdinov R.R., Vakhitov V.A. How to exclude the appearance of false-positive results during the polymerase chain reaction? Vestn. biotechnol. fiz.-chem. biol. 2012. V. 8(3). P. 34-45. (In Russian)

4. Chemeris D.A., Kiryanova O.Yu., Gubaydullin I.M., Chemeris A.V. Design of primers for polymerase chain reaction (Brief review of software and databases). Biomics. 2016. V. 8(3). P.215-238. (In Russian)

5. Chemeris D.A., Magdanov E.G., Mashkov O.I., Garafutdinov R.R., Chemeris A.V. Hot start or timerelease PCR. Biomics. 2011. V.2(1). P.1-8. (In Russian)

6. Garafutdinov R.R., Baymiev An.Kh., Maleev G.V., Alexeyev Ya.I., Zubov V.V., Chemeris D.A., Kiryanova J.Yu., Gubaydullin I.M., Matniyazov R.T., Sakhabutdinova A.R., Nikonorov Yu.M., Kuluev B.R., Baymiev Al.Kh., Chemeris A.V. Diversity of PCR primers and principles of their design. Biomics. 2019. V.11(1). P. 23 – 70. DOI: 10.31301/2221-6197.bmcs.2019-04 (In Russian)

7. Garafutdinov R.R., Galimova А.А., Sakhabutdinova А.R. Polymerase chain reaction with nearby primers // Anal. Biochem. 2017. V.518. P. 126-133. doi: 10.1016/j.ab.2016.11.017

8. Garafutdinov R.R., Galimova А.А., Sakhabutdinova А.R. The influence of quality of primers on the formation of primer dimers in PCR. Nucleos. Nucleot. Nucleic acids. 2020. V. 39(9). P. 1251-1269. doi: 10.1080/15257770.2020.1803354

9. Garafutdinov R.R., Nagaev N.R., Sakhabutdinova A.R., Chemeris A.V. Authenticity, integrity and availability of ancient DNA. Vestnik Bashkirskogo unversiteta. 2015. V.20(2). P. 432-439. (In Russian)

10. Karunanathie H, Kee PS, Ng SF, Kennedy MA, Chua EW. PCR enhancers: Types, mechanisms, and applications in long-range PCR. Biochimie. 2022. V.197. P.130-143. doi: 10.1016/j.biochi.2022.02.009

11. Ogata N, Miura T. Creation of genetic information by DNA polymerase of the thermophilic bacterium Thermus thermophilus // Nucleic Acids Res. 1998. V.26(20). P.4657-4661. doi: 10.1093/nar/26.20.4657

12. Yuce M., Kurt H., Mokkapati V.R.S.S., Budak B. Employment of nanomaterials in polymerase chain reaction: insight into the impacts and putative operating mechanisms of nano-additives in PCR. RSC Advances. 2014. V.4(69). P.36800-36814. doi: 10.1039/c4ra06144f 

Download pdf
up
eISSN: 2221-6197 DOI: 10.31301/2221-6197