Development of an easy and cost-effective method for non-invasive genotyping of insects

• DOI: 10.1371/journal.pone.0216998
• Publication Date: 2019-06-03T17:26:36Z
• Authors: Bahar Ali; Yicheng Zhou; Qiuyuan Zhang; Changying Niu; Zhihui Zhu
• Keywords: Frass

Non-invasive genotyping methods provide valuable information on insect populations. However, poor DNA amplification and time-consuming sampling procedures limit these methods, especially for small insects. An efficient convenient method was developed non-invasive, non-lethal of a large insect, Mythimna separata, Drosophila melanogaster, by endogenous exogenous, nuclear mitochondrial genes from frass, exuviae, food waste. For M. the chitin synthesis gene MsCHSB COI were successfully detected PCR exuviae DNA. fragment could not be directly probably due to degradation. To improve detection rate, frass first amplified Multiple Displacement Amplification with phi29 polymerase, after which all samples PCR. D. second instar larvae reared individually three days then extracted waste each individual. The serendipity α (sryα), exogenous transgene ΦC31 integrase, kl-5 gene, Y-chromosome-located male-specific marker most samples. We simple, determine gender identify transgenic individuals early in larval stage. This universal is applicable insects has potential application genetic ecological studies other arthropods.