A cost-effective, simple and high-throughput method for DNA extraction from insects

被引:14
作者
Lagisz, Malgorzata [1 ]
Port, Gordon [1 ]
Wolff, Kirsten [1 ]
机构
[1] Univ Newcastle, Sch Biol, Newcastle Upon Tyne NE1 7RU, Tyne & Wear, England
关键词
DNA; extraction; genetic; isolation; molecular; technique; PROTOCOL; AMPLIFICATION; PCR;
D O I
10.1111/j.1744-7917.2010.01318.x
中图分类号
Q96 [昆虫学];
学科分类号
摘要
We present a high-throughput cost-effective method to extract DNA suitable for polymerase chain reaction (PCR) from insect tissue. The method uses standard 200 mu L-deep 96-well plates in which samples are ground, digested and subsequently purified. The test extraction using four different insect species and controlling for potential contamination showed that the method yields good-quantity and quality DNA. PCR with mitochondrial and nuclear primers was reliable. The proposed extraction protocol combines the speed of commercial 96-well plate methods with the economies associated with readily available and cheap laboratory chemicals, consumables and equipment. Therefore, this method is particularly suitable for low-budget research projects and for laboratories with only basic equipment present.
引用
收藏
页码:465 / 470
页数:6
相关论文
共 14 条
  • [1] Ball SL, 2008, J ECON ENTOMOL, V101, P523, DOI 10.1603/0022-0493(2008)101[523:RODEFI]2.0.CO
  • [2] 2
  • [3] Folmer O., 1994, Molecular Marine Biology and Biotechnology, V3, P294
  • [4] A fast and inexpensive DNA extraction/purification protocol for brown macroalgae
    Hoarau, Galice
    Coyer, James A.
    Stam, Wytze T.
    Olsen, Jeanine L.
    [J]. MOLECULAR ECOLOGY NOTES, 2007, 7 (02): : 191 - 193
  • [5] Hoy M.A., 2003, Insect Molecular Genetics, V2nd
  • [6] Nondestructive DNA extraction from blackflies (Diptera: Simuliidae):: retaining voucher specimens for DNA barcoding projects
    Hunter, Stephanie J.
    Goodall, Tim I.
    Walsh, Kerry A.
    Owen, Richard
    Day, John C.
    [J]. MOLECULAR ECOLOGY RESOURCES, 2008, 8 (01) : 56 - 61
  • [7] An inexpensive, automation-friendly protocol for recovering high-quality DNA
    Ivanova, Natalia V.
    Dewaard, Jeremy R.
    Hebert, Paul D. N.
    [J]. MOLECULAR ECOLOGY NOTES, 2006, 6 (04): : 998 - 1002
  • [8] Amplification facilitators and multiplex PCR: Tools to overcome PCR-inhibition in DNA-gut-content analysis of soil-living invertebrates
    Juen, Anita
    Traugott, Michael
    [J]. SOIL BIOLOGY & BIOCHEMISTRY, 2006, 38 (07) : 1872 - 1879
  • [9] A SIMPLE SALTING OUT PROCEDURE FOR EXTRACTING DNA FROM HUMAN NUCLEATED CELLS
    MILLER, SA
    DYKES, DD
    POLESKY, HF
    [J]. NUCLEIC ACIDS RESEARCH, 1988, 16 (03) : 1215 - 1215
  • [10] Preparation and purification of DNA from insects for AFLP analysis
    Reineke, A
    Karlovsky, P
    Zebitz, CPW
    [J]. INSECT MOLECULAR BIOLOGY, 1998, 7 (01) : 95 - 99