Extraction of genomic DNA from Melipona quadrifasciata (Hymenoptera: Apidae, Meliponinae)

Abstract

The objective of the present study was to test three different procedures for DNA extraction of Melipona quadrifasciata based on existing methods for DNA extraction of Apis, plants and fungi. These methods differ in the concentrations of specific substances in the extraction buffer. The results demonstrate that the method used for Apis is not adequate for DNA extraction from M. quadrifasciata. On the other hand, with minor modifications this method and the methods for plants and fungi were adequate for DNA extraction of this stingless bee, both for adults and larvae<br>O objetivo deste trabalho foi o de testar três protocolos de extração de DNA utilizados para Apis, plantas e fungos, visando determinar um que seja eficiente para extração de DNA de Melipona quadrifasciata. Esses métodos diferem nas concentrações de componentes específicos do tampão de extração. Os resultados obtidos mostraram que a metodologia recomendada para extração de DNA de Apis não é adequada para a extração de DNA de M. quadrifasciata. Entretanto, com pequenas modificações, este método, bem como aquele utilizado para a extração de DNA de plantas e fungos, mostrou-se eficiente para a extração de DNA desta abelha sem ferrão, tanto de indivíduos adultos como de larva

Genetic divergence between populations of the stingless bee uruçu amarela (Melipona rufiventris group, Hymenoptera, Meliponini): is there a new Melipona species in the Brazilian state of Minas Gerais?

“…The protocol described by Waldschmidt et al (1997) was used to extract DNA from five bees per colony for 57 colonies, the genetic material from each bee being analyzed separately. Nine microsatellite loci (Mbi32 (GenBank AF002827), Mbi215 (AF002837), Mbi218 (AF002838), Mbi232 (AF002841), Mbi233 (AF002842), Mbi254 (AF002843), Mbi256 (AF002844), Mbi259 (AF 002845) and Mbi278 (AF002846)) were analyzed using the amplifications conditions described by Peters et al (1998) for Melipona bicolor.…”

Genetic variability of native populations of Wasmannia Forel (Hymenoptera: Formicidae) and their biogeographical implications

“…The genomic DNA of fi ve individuals from each nest was individually extracted according to Waldschmidt et al (1997) and amplifi ed by PCR-RAPD according to Williams et al (1990), with some modifi cations. The reaction mixture (25 l) consisted of 3 l DNA, 2.5 l enzyme buffer, 1.0 l triphosphate deoxyribonucleotide mixture (dATP, dTTP, dCTP, dGTP), 2.5 l primer, 4 M, 15.8 l miliQ water and 0.2 l Taq polymerase.…”

An optimized method for the generation of AFLP markers in a stingless bee (Melipona quadrifasciata) reveals a high degree of intracolonial genetic polymorphism

“…We initially tested a modified phenol/chloroform extraction that includes a proteinase K digestion step and addition of SDS. This method has been tested for genomic DNA analyses of stingless bees (Waldschmidt et al, 1997). Next, we tested an extraction protocol that uses a high salt (6 M NaCl)/ethanol precipitation of DNA in combination with SDS and a proteinase K digestion step.…”

“…Generally, these methods use CTAB and a phenol/chloroform extraction protocol. We modified this because previous studies describing DNA extraction procedures for stingless bees have shown the importance of a step incorporating proteinase K digestion and the substitution of CTAB by SDS (Waldschmidt et al, 1997) before the phenol/chloroform extraction. In addition, we tested a high-salt DNA extraction procedure that is frequently employed in microsatellite studies on bees (Paxton et al, 1996).…”