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Measuring the process and rate of exogenous DNA degradation during digestion in mice

This study aimed to perform qualitative and quantitative examination of DNA degradation during the digestion process in the mouse gut through PCR, qPCR and short tandem repeat (STR) analysis. Human blood leukocytes were gavaged into the digestive tract in mice. GAPDH, TH01, TPOX and D7S820 genes in...

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Autores principales: Xing, Ruiqi, Liu, Hui, Qi, Xia, Pan, Lingzi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9018913/
https://www.ncbi.nlm.nih.gov/pubmed/35440601
http://dx.doi.org/10.1038/s41598-022-10340-7
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author Xing, Ruiqi
Liu, Hui
Qi, Xia
Pan, Lingzi
author_facet Xing, Ruiqi
Liu, Hui
Qi, Xia
Pan, Lingzi
author_sort Xing, Ruiqi
collection PubMed
description This study aimed to perform qualitative and quantitative examination of DNA degradation during the digestion process in the mouse gut through PCR, qPCR and short tandem repeat (STR) analysis. Human blood leukocytes were gavaged into the digestive tract in mice. GAPDH, TH01, TPOX and D7S820 genes in the contents of the stomach and small intestine were analyzed with PCR and qPCR at various times pre- and post-gavage. Through STR analysis, 21 human genomic DNA loci were analyzed. The half-life of DNA degradation, and the relationship between the average peak area and digestion time were determined. The PCR results showed bands of amplified genes at pre-gavage (0 min) and post-gavage (40, 80 and 120 min) from the mouse stomach contents, whereas no DNA bands from small intestinal chyme were observed after gavage. The qPCR results revealed a significant decrease in DNA concentrations during 40–120 min in the mouse stomach after gavage. At 120 min, 85.62 ± 8.10% of the DNA was degraded, and the half-life of exogenous DNA degradation in the mouse stomach was 70.50 ± 5.46 min. At various digestion times, almost no target genes were detected in the mouse small intestinal chyme. STR analysis showed a decrease in allele numbers with bowel advancement in the small intestine in mice. The degradation of exogenous DNA was higher in the mouse stomach during the first 2 h, and almost complete degradation was observed within 40 min after entering the small intestine in mice.
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spelling pubmed-90189132022-04-21 Measuring the process and rate of exogenous DNA degradation during digestion in mice Xing, Ruiqi Liu, Hui Qi, Xia Pan, Lingzi Sci Rep Article This study aimed to perform qualitative and quantitative examination of DNA degradation during the digestion process in the mouse gut through PCR, qPCR and short tandem repeat (STR) analysis. Human blood leukocytes were gavaged into the digestive tract in mice. GAPDH, TH01, TPOX and D7S820 genes in the contents of the stomach and small intestine were analyzed with PCR and qPCR at various times pre- and post-gavage. Through STR analysis, 21 human genomic DNA loci were analyzed. The half-life of DNA degradation, and the relationship between the average peak area and digestion time were determined. The PCR results showed bands of amplified genes at pre-gavage (0 min) and post-gavage (40, 80 and 120 min) from the mouse stomach contents, whereas no DNA bands from small intestinal chyme were observed after gavage. The qPCR results revealed a significant decrease in DNA concentrations during 40–120 min in the mouse stomach after gavage. At 120 min, 85.62 ± 8.10% of the DNA was degraded, and the half-life of exogenous DNA degradation in the mouse stomach was 70.50 ± 5.46 min. At various digestion times, almost no target genes were detected in the mouse small intestinal chyme. STR analysis showed a decrease in allele numbers with bowel advancement in the small intestine in mice. The degradation of exogenous DNA was higher in the mouse stomach during the first 2 h, and almost complete degradation was observed within 40 min after entering the small intestine in mice. Nature Publishing Group UK 2022-04-19 /pmc/articles/PMC9018913/ /pubmed/35440601 http://dx.doi.org/10.1038/s41598-022-10340-7 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Xing, Ruiqi
Liu, Hui
Qi, Xia
Pan, Lingzi
Measuring the process and rate of exogenous DNA degradation during digestion in mice
title Measuring the process and rate of exogenous DNA degradation during digestion in mice
title_full Measuring the process and rate of exogenous DNA degradation during digestion in mice
title_fullStr Measuring the process and rate of exogenous DNA degradation during digestion in mice
title_full_unstemmed Measuring the process and rate of exogenous DNA degradation during digestion in mice
title_short Measuring the process and rate of exogenous DNA degradation during digestion in mice
title_sort measuring the process and rate of exogenous dna degradation during digestion in mice
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9018913/
https://www.ncbi.nlm.nih.gov/pubmed/35440601
http://dx.doi.org/10.1038/s41598-022-10340-7
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