Advance Search

Message Board

Respected readers, authors and reviewers, you can add comments to this page on any questions about the contribution, review, editing and publication of this journal. We will give you an answer as soon as possible. Thank you for your support!

Name
E-mail
Phone
Title
Content
Verification Code
Volume 8 Issue 3
Sep.  2017
Turn off MathJax
Article Contents
Article Navigation >  Journal of Tropical Biology  > 2017  >  8(3): 267-276

DING Qing, LIU Yuepeng, ZHU Xiaopeng, WU Kebang, LUO Sulan, ZHANGSUN Dongting. Separation and Preparation of Different Size Genomic DNA Fragments of Conus Vexillum[J]. Journal of Tropical Biology, 2017, 8(3): 267-276. doi: 10.15886/j.cnki.rdswxb.2017.03.004
Citation: DING Qing, LIU Yuepeng, ZHU Xiaopeng, WU Kebang, LUO Sulan, ZHANGSUN Dongting. Separation and Preparation of Different Size Genomic DNA Fragments of Conus Vexillum[J]. Journal of Tropical Biology, 2017, 8(3): 267-276. doi: 10.15886/j.cnki.rdswxb.2017.03.004
shu

Separation and Preparation of Different Size Genomic DNA Fragments of Conus Vexillum

doi: 10.15886/j.cnki.rdswxb.2017.03.004

  • 1. Key Laboratory of Tropical Biological Resources of Ministry of Education, Key Laboratory for Marine Drug of Haikou, Hainan University, Haikou 570228, China;

  • 2. College of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China

  • Received Date: 2017-03-13
  • Rev Recd Date: 2017-04-17
  • In order to provide basis for construction of the genomic library of Conus vexillum（C.vexillum）,different methods were used for isolation and extraction of different size fragments of genomic DNA from C.vexillum native to Hainan.Genomic DNA was isolated from venom glands,hepatopancreas and muscle of C.vexillum using three methods of CTAB,spin column and magnetic bead respectively.The genomic DNA fragments were separated by agarose gel electrophoresis and pulsed-field gel electrophoresis,of which the size and distribution range of extracted DNA was detected.The results showed that the genomic DNA could be extracted by all the three methods.The purity and yield of DNAs isolated by CTAB and centrifugal column methods using venom gland of C.vexillum were good and high,which could meet the following experimental requirements.While the yield of the magnetic bead method is less and the purity is not good.The length of most DNA fragments isolated by the centrifugal column method was less than 9 kb,while most fragments’ sizes isolated by CTAB method were more than 20 kb.Among the three kinds of organs,hepatopancreas had highest yield of DNA,but its fragment was dispersed with degradation happened,the DNA yield of venom glands was high but less than hepatopancreas slightly,its integrity was the best;however the DNA yield of muscle was lowest and its fragments was smaller.The high quality genomic DNA fragments with different sizes were separated and prepared successfully by optimized pulsed-field gel electrophoresis.In conclusion,using CTAB method to isolate C.vexillum DNA is better than the other methods for big fragment genomic DNA library construction.Different size genomic DNA fragments recovered by pulsed-field gel electrophoresis would lay a foundation for further genomic library construction of C.vexillum using different gene library vector systems.
  • 加载中
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索
Get Citation
PDF
XML

Article views(2) PDF downloads(0) Cited by()

Proportional views
Related

Separation and Preparation of Different Size Genomic DNA Fragments of Conus Vexillum

doi: 10.15886/j.cnki.rdswxb.2017.03.004
  • 1. Key Laboratory of Tropical Biological Resources of Ministry of Education, Key Laboratory for Marine Drug of Haikou, Hainan University, Haikou 570228, China;
  • 2. College of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China
  • Received Date: 2017-03-13
  • Rev Recd Date: 2017-04-17

Abstract: In order to provide basis for construction of the genomic library of Conus vexillum（C.vexillum）,different methods were used for isolation and extraction of different size fragments of genomic DNA from C.vexillum native to Hainan.Genomic DNA was isolated from venom glands,hepatopancreas and muscle of C.vexillum using three methods of CTAB,spin column and magnetic bead respectively.The genomic DNA fragments were separated by agarose gel electrophoresis and pulsed-field gel electrophoresis,of which the size and distribution range of extracted DNA was detected.The results showed that the genomic DNA could be extracted by all the three methods.The purity and yield of DNAs isolated by CTAB and centrifugal column methods using venom gland of C.vexillum were good and high,which could meet the following experimental requirements.While the yield of the magnetic bead method is less and the purity is not good.The length of most DNA fragments isolated by the centrifugal column method was less than 9 kb,while most fragments’ sizes isolated by CTAB method were more than 20 kb.Among the three kinds of organs,hepatopancreas had highest yield of DNA,but its fragment was dispersed with degradation happened,the DNA yield of venom glands was high but less than hepatopancreas slightly,its integrity was the best;however the DNA yield of muscle was lowest and its fragments was smaller.The high quality genomic DNA fragments with different sizes were separated and prepared successfully by optimized pulsed-field gel electrophoresis.In conclusion,using CTAB method to isolate C.vexillum DNA is better than the other methods for big fragment genomic DNA library construction.Different size genomic DNA fragments recovered by pulsed-field gel electrophoresis would lay a foundation for further genomic library construction of C.vexillum using different gene library vector systems.

DING Qing, LIU Yuepeng, ZHU Xiaopeng, WU Kebang, LUO Sulan, ZHANGSUN Dongting. Separation and Preparation of Different Size Genomic DNA Fragments of Conus Vexillum[J]. Journal of Tropical Biology, 2017, 8(3): 267-276. doi: 10.15886/j.cnki.rdswxb.2017.03.004
Citation: DING Qing, LIU Yuepeng, ZHU Xiaopeng, WU Kebang, LUO Sulan, ZHANGSUN Dongting. Separation and Preparation of Different Size Genomic DNA Fragments of Conus Vexillum[J]. Journal of Tropical Biology, 2017, 8(3): 267-276. doi: 10.15886/j.cnki.rdswxb.2017.03.004
shu

    HTML

Catalog

    Copyright © 《Journal of Tropical Biology》Editorial Board

    Address: 58 Renmin Avenue, Haikou, Hainan China Pos: 100101Tel: 0898-66281595

    Supported by: Beijing Renhe Information Technology Co. Ltd

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return