Genomic DNA extraction from honey bee ( Apis mellifera ) queen spermathecal content

AbstractGenetic analysis of the honey bee spermathecal content can be particularly useful to provide an estimate of the genetic diversity and purity of the surrounding populations. Here we compared the concentration and quality of DNA extracted from queen spermatheca using four commercial kits to determine the best method to obtain DNA suitable for single nucleotide polymorphism genotyping by next-generation sequencing. The four kits were tested with different adjustments in the lysis incubation time, use of RNA-carrier, elution conditions and number of re-elutions. Only the use of QIAamp DNA Microkit with 3 h of lysis incubation, the addition of RNA-carrier and multiple re-elutions produced a DNA concentration over the required threshold.Keywords: Honey beespermathecagenomic DNA isolationDNA extraction Disclosure statementNo potential conflict of interest was reported by the authors.Additional informationFundingContributions of Jakob Wegener and Eduard Musin were supported by funds of the Federal Ministry of Food and Agriculture based on a decision of the Parliament of the Federal Republic of Germany via the Federal Office for Agriculture and Food, grant number 2818BM040. Dora Henriques was supported by BEEHAPPY (POCI-01-0145-FEDER-029871; FCT and COMPETE/QREN/EU). Ana R. Lopes and Carlos A. Yadró were supported by Fundação para a Ciência e a Tecnologia (FCT) grants SFRH/BD/143627/2019 and 2021.06948.BD, respectively. FCT provided financial support by national funds (FCT/MCTES) to CIMO (UIDB/00690/2020 and UIDP/00690/2020) and SusTEC (LA/P/0007/2021).