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異なるDNA精製法が定量PCRによるEdwardsiella tardaの計数に与える影響


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Title: 異なるDNA精製法が定量PCRによるEdwardsiella tardaの計数に与える影響
Other Titles: Effect of different DNA purification kits on counting the cell number of Edwardsiella tarda using quantitative PCR
Authors: 朝重, 紅音 / 菅, 向志郎 / 金井, 欣也
Authors (alternative): Tomoshige, Akane / Suga, Koushirou / Kanai, Kinya
Issue Date: Mar-2018
Publisher: 長崎大学水産学部 / The Faculty of Fisheries, Nagasaki University
Citation: 長崎大学水産学部研究報告, 99, pp.7-11; 2018
Abstract: Quantitative PCR (qPCR) quantifies the template DNA from the number of threshold cycles (Ct value), but the probability of quantitative errors occurs due to the differences in recovery rates during DNA purification. In order to investigate the effect of different DNA purification kits in counting Edwardsiella tarda cells using qPCR, we compared two methods; the bacterial culture dilution method, in which the DNA is purified after 5 steps of a ten-fold serial dilution of E. tarda culture, and the DNA dilution method, which the DNA is diluted after purification with the optimum cell number of E. tarda. In both methods, we used three types of DNA purification kit: Chelex resin, Non-Silica membrane and Silica membrane. Our results showed the lowest Ct value and high DNA recovery rate were obtained using Chelex resin. Non-Silica membrane and Silica membrane Ct values in DNA dilution method were higher than that of Chelex resin. With the use of Non-Silica membrane, the calculated DNA using bacterial culture dilution method decreased by 67-87% compared to DNA dilution method at 1.0×10 4cells/μL of E. tarda.
Keywords: 定量PCR / 細菌計数 / DNA精製 / Edwardsiella tarda / quantitative PCR / bacterial count / DNA purification
URI: http://hdl.handle.net/10069/37991
ISSN: 05471427
Type: Departmental Bulletin Paper
Text Version: publisher
Appears in Collections:No. 99

Citable URI : http://hdl.handle.net/10069/37991

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