Pretreatment Method for DNA Barcoding to Analyze Gut Contents of Rotifers
Autor: | Sun-Jin Hwang, Jeong-Soo Gim, Hye-Ji Oh, Gea-Jae Joo, Kwang-Hyeon Chang, Hyun-Gi Jeong, Hyunbin Jo, Ihn-Sil Kwak, Paul Henning Krogh |
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Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
0106 biological sciences
Cyanobacteria Bleach eliminate the extracellular DNA Chlorophyceae Rotifer 010603 evolutionary biology 01 natural sciences DNA barcoding lcsh:Technology lcsh:Chemistry commercial bleach chemistry.chemical_compound General Materials Science Food science Instrumentation lcsh:QH301-705.5 Fluid Flow and Transfer Processes biology lcsh:T 010604 marine biology & hydrobiology Process Chemistry and Technology General Engineering pretreatment biology.organism_classification lcsh:QC1-999 Computer Science Applications chemistry lcsh:Biology (General) lcsh:QD1-999 lcsh:TA1-2040 gut content of Rotifera Lorica (biology) lcsh:Engineering (General). Civil engineering (General) DNA Bacteria lcsh:Physics |
Zdroj: | Applied Sciences Volume 10 Issue 3 Applied Sciences, Vol 10, Iss 3, p 1064 (2020) |
ISSN: | 2076-3417 |
DOI: | 10.3390/app10031064 |
Popis: | We designed an experiment to analyze the gut content of Rotifera based on DNA barcoding and tested it on Asplanchna sp. in order to ensure that the DNA extracted from the rotifer species is from the food sources within the gut. We selected ethanol fixation (60%) to minimize the inflow effects of treated chemicals, and commercial bleach (the final concentration of 2.5%, for 210 s) to eliminate the extracellular DNA without damage to the lorica. Rotifers have different lorica structures and thicknesses. Therefore, we chose a pretreatment method based on Asplanchna sp., which is known to have weak durability. When we used the determined method on a reservoir water sample, we confirmed that the DNA fragments of Chlorophyceae, Diatomea, Cyanobacteria, and Ciliophora were removed. Given this result, Diatomea and cyanobacteria, detected from Asplanchna, can be considered as gut contents. However, bacteria were not removed by bleach, thus there was still insufficient information. Since the results of applying commercial bleach to rotifer species confirmed that pretreatment worked effectively for some species of rotifers food sources, in further studies, it is believed to be applicable to the gut contents analysis of more diverse rotifers species and better DNA analysis techniques by supplementing more rigorous limitations. |
Databáze: | OpenAIRE |
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