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作者:

Gao, Jingfeng (Gao, Jingfeng.) (学者:高景峰) | Pan, Kailing (Pan, Kailing.) | Li, Hongyu (Li, Hongyu.) | Fan, Xiaoyan (Fan, Xiaoyan.) (学者:樊晓燕) | Sun, Lixin (Sun, Lixin.) | Zhang, Shujun (Zhang, Shujun.) | Gao, Yongqing (Gao, Yongqing.)

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摘要:

In this study, GelGreen (TM) was investigated as a replacement for SYBR (R) Safe to stain DNA in cesium chloride (CsCI) density gradients for DNA-stable isotope probing (SIP) experiments. Using environmental DNA, the usage of GelGreen (TM) was optimized for sensitivity compared to SYBR (R) Safe, its optimal concentration, detection limit for environmental DNA and its application in environmental DNA-SIP assay. Results showed that GelGreen (TM) was more sensitive than SYBR (R) Safe, while the optimal dosage (15X concentration) needed was approximately one-third of SYBR (R) Safe, suggesting that its sensitivity was three times more superior than SYBR (R) Safe. At these optimal parameters, the detection limit of Gel-Green (TM)-stained environmental DNA was as low as 0.2 mu g, but the usage of 0.5 mu g environmental DNA was recommended to produce a more consistent DNA band. In addition, a modified needle extraction procedure was developed to withdraw DNA effectively by fractionating CsCI density gradients into four or five fractions. The successful application of Gel-Green (TM) staining with C-13-labeled DNA from enriched activated sludge suggests that this stain was an excellent alternative of SYBR (R) Safe in CsCI density gradients for DNA-SIP assays.

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作者机构:

  • [ 1 ] [Gao, Jingfeng]Beijing Univ Technol, Coll Environm & Energy Engn, Natl Engn Lab Adv Municipal Wastewater Treatment, Beijing, Peoples R China
  • [ 2 ] [Pan, Kailing]Beijing Univ Technol, Coll Environm & Energy Engn, Natl Engn Lab Adv Municipal Wastewater Treatment, Beijing, Peoples R China
  • [ 3 ] [Li, Hongyu]Beijing Univ Technol, Coll Environm & Energy Engn, Natl Engn Lab Adv Municipal Wastewater Treatment, Beijing, Peoples R China
  • [ 4 ] [Fan, Xiaoyan]Beijing Univ Technol, Coll Environm & Energy Engn, Natl Engn Lab Adv Municipal Wastewater Treatment, Beijing, Peoples R China
  • [ 5 ] [Sun, Lixin]Beijing Univ Technol, Coll Environm & Energy Engn, Natl Engn Lab Adv Municipal Wastewater Treatment, Beijing, Peoples R China
  • [ 6 ] [Zhang, Shujun]Beijing Drainage Grp Co Ltd, Ctr Res & Dev, Beijing, Peoples R China
  • [ 7 ] [Gao, Yongqing]Beijing Drainage Grp Co Ltd, Ctr Res & Dev, Beijing, Peoples R China

通讯作者信息:

  • 高景峰

    [Gao, Jingfeng]Beijing Univ Technol, Coll Environm & Energy Engn, Natl Engn Lab Adv Municipal Wastewater Treatment, Beijing, Peoples R China

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来源 :

PLOS ONE

ISSN: 1932-6203

年份: 2017

期: 1

卷: 12

3 . 7 0 0

JCR@2022

ESI学科: Multidisciplinary;

ESI高被引阀值:283

中科院分区:3

被引次数:

WoS核心集被引频次: 3

SCOPUS被引频次: 3

ESI高被引论文在榜: 0 展开所有

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