Comparison of preservation techniques for silkworm (Bombyx mori L.) DNA based on polymerase chain reaction (PCR) products
2003
Jiraporn Tayutivutikul(Chiang Mai University, Chiang Mai (Thailand). Faculty of Agriculture. Department of Entomology) E-mail:j.tayuti@chiangmai.ac.th | Weerathep Pongprasert(Naresuan University, Phitsanulok (Thailand). Faculty of Agriculture, Natural Resources and Environment. Department of Agricultural Science) | Royce, Lynn A.(Oregon State University, Corvallis (USA). Department of Entomology) | Krissana Ruangrit(Chiang Mai University, Chiang Mai (Thailand). Faculty of Agriculture. Department of Entomology)
There are many ways to preserve insect specimens that will protect its DNA from degradation for the period of time from collection to use in a molecular genetic study. However, techniques vary among groups of insects, suggesting that the determination of preservation should be done before working at the molecular level with large number of individuals from a particular group. This work compares four preservation techniques for the Thai native silkworm, Bombyx mori L. (UB1XNangnoi Srisaket 1): (1) killed and stored at -20 deg C, (2) killed and stored at -80 deg C, (3) killed and stored in 70 percent ethanol, and (4) rapid hot-air drying at 60 deg C. All specimens were stored for six months. DNA of each specimen was extracted by two different methods: (1) freezing in liquid nitrogen and transferring to lysis buffer for grinding and (2) grinding directly in lysis buffer. The integrity of DNA from those were determined and compared to live specimens using amplification of COI-COII DNA as a target region and genetic mapping with RAPDs. The method using 70 percent ethanol was found to be most practical and could prevent degradation of silkworm DNA for at least six months.
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