Freezing method as a new non-destructive modification of DNA extraction

Salmah Yaakop, C. Van Achterberg, Idris Abd. Ghani, A. Z. Aman

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A modification of a DNA extraction method by freezing specimens is recognized as one of new non-destructive techniques. In this study, the freezing method has been applied on dried and fresh, tiny and economically important insect samples, i.e. on adults and larvae of wasps, fruit flies and thrips. The modification entails freezing instead of a lengthy incubation of the sample. Most importantly, the sample is not cut into small pieces, but is soaked in a lysis buffer and then frozen in -22°C for a minimum of 20 minutes. After that, the remaining protocols from the manual of DNeasy Blood and Tissue Kit are followed. Several other non-destructive methods also require incubation for at least 20 minutes in a lysis buffer at 55°C. However, the duration of that incubation process is not standard for all insect and arthropod species. This is because the optimization process is based on species size and the thickness of the insect cuticle. With the freezing method, samples are not damaged, and remain available for morphological re-examination. Hence, the sample can also be re-used for taxonomic work with no distortion of samples, no loss of coloration and no phenotypic changes on the external morphology. The complete protocol for the freezing method is described in this paper. With this freezing method, DNA concentration of 0.2- 5.61 ng/μl was recovered on various tiny insect species. Furthermore. several specimens of Bactrocera and Heratemis species were selected as control specimens in analyzing a variety of extraction methods. The freezing method was proven as a new technique to obtain sufficient quantity and a high quality of DNA for molecular work.

Original languageEnglish
Pages (from-to)373-392
Number of pages20
JournalPertanika Journal of Tropical Agricultural Science
Volume36
Issue number4
Publication statusPublished - Nov 2013

Fingerprint

Freezing
freezing
DNA
Insects
insect
incubation
lysis
extraction method
sampling
insects
Buffers
buffers
Bactrocera
insect cuticle
Thysanoptera
Tephritidae
thrips
nondestructive methods
Wasps
cuticle

Keywords

  • Freezing method
  • Molecular DNA
  • Non-destructive samples
  • Phylogenetic analysis
  • Sufficient DNA

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Agricultural and Biological Sciences(all)
  • Chemical Engineering(all)
  • Computer Science(all)
  • Environmental Science(all)

Cite this

Freezing method as a new non-destructive modification of DNA extraction. / Yaakop, Salmah; Van Achterberg, C.; Abd. Ghani, Idris; Aman, A. Z.

In: Pertanika Journal of Tropical Agricultural Science, Vol. 36, No. 4, 11.2013, p. 373-392.

Research output: Contribution to journalArticle

Yaakop, Salmah ; Van Achterberg, C. ; Abd. Ghani, Idris ; Aman, A. Z. / Freezing method as a new non-destructive modification of DNA extraction. In: Pertanika Journal of Tropical Agricultural Science. 2013 ; Vol. 36, No. 4. pp. 373-392.
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