{"title":"利用大肠杆菌和火星沙模拟物进行聚合酶链反应实验,以探测和分析地外生命","authors":"Keigo Enya , Satoshi Sasaki , Taiki Kunieda","doi":"10.1016/j.lssr.2024.05.003","DOIUrl":null,"url":null,"abstract":"<div><p>In this study, we conducted polymerase chain reaction (PCR) experiments using <em>Escherichia coli</em> (<em>E. coli</em>) and a Mars sand simulant (Mars Global Simulant MGS-1, Exolith Lab) to detect and analyze potential extraterrestrial life. The targeted DNA sequence is common among the bacterial kingdom on Earth. PCR experiments conducted after alkaline heat extraction, wherein samples with varying amounts of Mars sand simulant were compared, revealed that the simulant interfered with DNA detection. We then conducted PCR experiments following treatment with a sand DNA extraction kit on samples with various <em>E. coli</em> densities. DNA bands for a minimum <em>E. coli</em> density of 900 cells/(g sand) were confirmed, while no DNA bands were visible in the 90 cells/(g sand) sample with and without the Mars sand simulant. The total DNA mass contained in 900 cells was calculated to be 15.3 pg (i.e., 1.53 pg in 0.1 g sand sample we evaluated). We tested and compared the influence of the eluate of Mars sand simulant and DNA adsorption onto Mars sand simulant based on optical absorbance measurements. Our findings suggest that the mechanism by which the Mars sand simulant prevents PCR is through the adsorption of DNA onto the Mars sand simulant.</p></div>","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":null,"pages":null},"PeriodicalIF":4.6000,"publicationDate":"2024-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214552424000610/pdfft?md5=636150b80cc1f7b27f024fbfc6c2710a&pid=1-s2.0-S2214552424000610-main.pdf","citationCount":"0","resultStr":"{\"title\":\"A polymerase chain reaction experiment using Escherichia coli and Mars sand simulant for detection and analysis of extraterrestrial life\",\"authors\":\"Keigo Enya , Satoshi Sasaki , Taiki Kunieda\",\"doi\":\"10.1016/j.lssr.2024.05.003\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>In this study, we conducted polymerase chain reaction (PCR) experiments using <em>Escherichia coli</em> (<em>E. coli</em>) and a Mars sand simulant (Mars Global Simulant MGS-1, Exolith Lab) to detect and analyze potential extraterrestrial life. The targeted DNA sequence is common among the bacterial kingdom on Earth. PCR experiments conducted after alkaline heat extraction, wherein samples with varying amounts of Mars sand simulant were compared, revealed that the simulant interfered with DNA detection. We then conducted PCR experiments following treatment with a sand DNA extraction kit on samples with various <em>E. coli</em> densities. DNA bands for a minimum <em>E. coli</em> density of 900 cells/(g sand) were confirmed, while no DNA bands were visible in the 90 cells/(g sand) sample with and without the Mars sand simulant. The total DNA mass contained in 900 cells was calculated to be 15.3 pg (i.e., 1.53 pg in 0.1 g sand sample we evaluated). We tested and compared the influence of the eluate of Mars sand simulant and DNA adsorption onto Mars sand simulant based on optical absorbance measurements. Our findings suggest that the mechanism by which the Mars sand simulant prevents PCR is through the adsorption of DNA onto the Mars sand simulant.</p></div>\",\"PeriodicalId\":2,\"journal\":{\"name\":\"ACS Applied Bio Materials\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.6000,\"publicationDate\":\"2024-05-23\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2214552424000610/pdfft?md5=636150b80cc1f7b27f024fbfc6c2710a&pid=1-s2.0-S2214552424000610-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"ACS Applied Bio Materials\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2214552424000610\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MATERIALS SCIENCE, BIOMATERIALS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Applied Bio Materials","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2214552424000610","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MATERIALS SCIENCE, BIOMATERIALS","Score":null,"Total":0}
引用次数: 0
摘要
在这项研究中,我们利用大肠杆菌(E. coli)和火星沙模拟物(火星全球模拟物 MGS-1,Exolith Lab)进行了聚合酶链反应(PCR)实验,以探测和分析潜在的地外生命。目标 DNA 序列在地球细菌王国中很常见。在碱热提取后进行的 PCR 实验中,我们对含有不同量火星沙模拟物的样品进行了比较,结果发现模拟物会干扰 DNA 检测。随后,我们使用沙粒 DNA 提取试剂盒对不同大肠杆菌密度的样本进行了 PCR 实验。结果证实,大肠杆菌最小密度为 900 个细胞/(克沙)时,DNA 条带清晰可见,而 90 个细胞/(克沙)的样本在使用或未使用火星沙模拟剂时,DNA 条带均不清晰可见。经计算,900 个细胞所含的 DNA 总量为 15.3 pg(即我们评估的 0.1 g 沙样本中含有 1.53 pg)。我们测试并比较了火星沙模拟物洗脱液和 DNA 吸附在火星沙模拟物上的影响(基于光学吸光度测量)。我们的研究结果表明,火星沙模拟物阻止 PCR 的机制是通过 DNA 在火星沙模拟物上的吸附。
A polymerase chain reaction experiment using Escherichia coli and Mars sand simulant for detection and analysis of extraterrestrial life
In this study, we conducted polymerase chain reaction (PCR) experiments using Escherichia coli (E. coli) and a Mars sand simulant (Mars Global Simulant MGS-1, Exolith Lab) to detect and analyze potential extraterrestrial life. The targeted DNA sequence is common among the bacterial kingdom on Earth. PCR experiments conducted after alkaline heat extraction, wherein samples with varying amounts of Mars sand simulant were compared, revealed that the simulant interfered with DNA detection. We then conducted PCR experiments following treatment with a sand DNA extraction kit on samples with various E. coli densities. DNA bands for a minimum E. coli density of 900 cells/(g sand) were confirmed, while no DNA bands were visible in the 90 cells/(g sand) sample with and without the Mars sand simulant. The total DNA mass contained in 900 cells was calculated to be 15.3 pg (i.e., 1.53 pg in 0.1 g sand sample we evaluated). We tested and compared the influence of the eluate of Mars sand simulant and DNA adsorption onto Mars sand simulant based on optical absorbance measurements. Our findings suggest that the mechanism by which the Mars sand simulant prevents PCR is through the adsorption of DNA onto the Mars sand simulant.