Comparison of DNA Quantity Extracted from Formalin and Ethanol Fixed Tissues

##plugins.themes.bootstrap3.article.main##

Piangjai Chalermwong Parinya Seelanan Patchara Sinloyma

Abstract

This study was performed to compare DNA quantity extracted from the cerebrum part of porcine brain tissues preserved in 10% formalin or 95% ethanol, for 8 weeks. A sample was collected and DNA extracted at least once a week using a QIAamp DNA FFPE Tissue Kit. The Qubit 4 Fluorometer was then used for DNA quantification and to study the association between preservation duration and fixatives to evaluate the effect of two factors on DNA quantity, which can support forensic officers in choosing the most effective fixative to preserve tissues for DNA testing. The results showed that the concentration of extracted DNA from the tissues preserved in 95% ethanol was significantly higher than in the 10% formalin-fixed tissues. Moreover, DNA concentration from tissues preserved in 10% formalin significantly decreased after one week of preservation, while no significant difference was observed between DNA concentration in 95% ethanol fixed tissues after preservation. In addition, the negative correlation between preservation duration and DNA quantity in both 10% formalin and 95% ethanol fixed tissues was observed. It can be described that extracted DNA concentrations from tissues preserved in 10% formalin and 95% ethanol were decreased when the preservation time increased. However, we found that DNA concentration from 10% formalin-fixed tissues reduced than tissues preserved in 95% ethanol by 40%. Therefore, it can be concluded that 95% ethanol has better preservation properties and can be an alternative for a suitable method to preserve DNA from brain tissues compared to the 10% formalin solution.


Keywords: DNA, ethanol, formalin, preservation

References

Akoglu, H. (2018). User's guide to correlation coefficients. Turkish Journal of Emergency Medicine, 18, 91–93.
http://dx.doi.org/10.1016/j.tjem.2018.08.001
Alex, E. (2020). Famous celebrities who had to get paternity tests. Retrieved from https://kiwireport.com/
famous-celebrities-who-had-to-get-paternity-tests/
Alqaydi, M., & Roy, R. (2016). Quantitative and qualitative study of STR DNA from ethanol and formalin fixed tissues. Forensic Science International, 262, 18-29. https://doi.org/10.1016/j.forsciint.2016.02.011
Douglas, M. P., & Rogers, S. O. (1998). DNA damage caused by common cytological fixatives. Mutat Res, 401(1-2), 77-88. https://doi.org/10.1016/S0027-5107(97)00314-X
Eltoum, I., Fredenburgh, J., Myers, R. B., & Grizzle, W. E. (2001). Introduction to the theory and practice of fixation of tissues. Journal of Histotechnology, 24(3), 173-190. https://doi.org/10.1179/his.2001.
24.3.173
Gino, S., Varacalli, S., Robino, C., & Torre, C. (2004). STR typing of fixed human tissue: formalin vs. an alcohol-based method. International Congress Series, 1261, 611-612. https://doi.org/10.1016/
S0531-5131(03)01660-1
Góes, A. C., Silva, D. A., Domingues, C. S., Sobrinho, J. M., & Carvalho, E. F. (2002). Identification of a criminal by DNA typing in a rape case in Rio de Janeiro, Brazil. Sao Paulo Medical Journal, 120(3), 77-79. https://doi.org/10.1590/S1516-31802002000300004
Howat, W. J., & Wilson, B. A. (2014). Tissue fixation and the effect of molecular fixatives on downstream staining procedures. Methods, 70(1), 12–19. https://doi.org/10.1016/j.ymeth.2014.01.022
Josefiova, J., Matura, R., Votrubova, J., Vojacek, T., Tomasek, P., & Vanek, D. (2017). Comparison of fluorometric and real- time PCR quantification of DNA extracted from formalin fixed tissue. Forensic Science International: Genetics Supplement Series, 6, e137–e139. https://doi.org/10.1016/j.fsigss.2017.09.028
Kumar, N., Maitray, A., Gupta, R., & Shukla, S. K. (2018). Effects of preservative on foetus tissues and DNA profiling in forensic cases. International Journal of Molecular Biology, 3(4), 165-167. https://doi.org/
10.15406/ijmboa.2018.03.00071
Lu, K., Ye, W., Zhou, L., Collins, L. B., Chen, X., Gold, A., … Swenberg, J. A. (2010). Structural characterization of formaldehyde-induced cross-links between amino acids and deoxynucleosides and their oligomers. Journal of the American Chemical Society, 132(10), 3388-3399. https://doi.org/10.1021/ja908282f
McGhee, J. D., & Von Hippel, P. H. (1975). Formaldehyde as a probe of DNA structure. I. Reaction with exocyclic amino groups of DNA bases. Biochemistry, 14(6), 1281-1296. https://doi.org/10.1021/bi00677a029
McGhee, J. D., & Von Hippel, P. H. (1975). Formaldehyde as a probe of DNA structure. II. Reaction with endocyclic imino groups of DNA bases. Biochemistry, 14(6), 1297-1303. https://doi.org/10.1021/bi00677a030
McGhee, J. D., & Von Hippel, P. H. (1977). Formaldehyde as a probe of DNA structure. 3. Equilibrium denaturation of DNA and synthetic Polynucleotides. Biochemistry, 16(15), 3267-3276. https://doi.
org/10.1021/bi00634a001
Marquina, D., Ronquist, F., & Lukasik, P. (2020). The effect of ethanol concentration on the preservation of insects for biodiversity studies. Retrieved from https://www.biorxiv.org/content/10.1101/2020.03.05.978288v2
Miething, F., Hering, S., Hanschke, B., & Dressler, J. (2016). Effect of fixation to the degradation of nuclear and mitochondrial DNA in different tissues. Journal of Histochemistry & Cytochemistry, 54(3), 371–374. https://doi.org/10.1369/jhc.5B6726.2005
Montelius, K., & Lindblom, B. (2012). DNA analysis in disaster victim identification. Forensic Science, Medicine and Pathology, 8, 140–147. https://doi.org/10.1007/s12024-011-9276-z
Paireder, S., Werner, B., Bailer, J., Werther, W., Schmid, E., Patzak, B., & Cichna-Markl, M. (2013). Comparison of protocols for DNA extraction from long-term preserved formalin fixed tissues. Analytical Biochemistry, 439, 152-160. https://doi.org/10.1016/j.ab.2013.04.006
Primorac, D., Andelinovic, S., Definis-Gojanovic, M., Drmic, I., Rezic, B., Baden, M. M., … Lee, H. C. (1996). Identification of war victims from mass graves in Croatia, Bosnia, and Herzegovina by use of standard forensic methods and DNA typing. Journal of forensic sciences, 41(5), 891-894. https://doi.
org/10.1520/JFS14019J.
Srinivasan, M., Sedmak, D., & Jewell, S. (2002). Effect of fixatives and tissue processing on the content and integrity of nucleic acids. American Journal of Pathology, 161(6), 1961-1971. https://doi.org/
10.1016/S0002-9440(10)64472-0
Usharani, M., Thilaga, D., & Mahalakshmi, N. (2019). A Comparative study on forensic tissue specimen preserved in formalin and sodium chloride. Forensic Research, 10(2), 1-5.
Werner, M., Chott, A., Fabiano, A., & Battifora, H. (2000). Effect of formalin tissue fixation and processing on immunohistochemistry. The American Journal of Surgical Pathology, 24(7), 1016–1019. https://doi.org/10.1097/00000478-200007000-00014

Section
Research Articles

##plugins.themes.bootstrap3.article.details##

How to Cite
CHALERMWONG, Piangjai; SEELANAN, Parinya; SINLOYMA, Patchara. Comparison of DNA Quantity Extracted from Formalin and Ethanol Fixed Tissues. Naresuan University Journal: Science and Technology (NUJST), [S.l.], v. 30, n. 4, p. 75-83, sep. 2022. ISSN 2539-553X. Available at: <https://www.journal.nu.ac.th/NUJST/article/view/Vol-30-No-4-2022-75-83>. Date accessed: 27 apr. 2024. doi: https://doi.org/10.14456/nujst.2022.37.