The Service Department offers a variety of high quality and rapid services to IMP and IMBA scientists. The majority of our effort involves DNA sequencing, fly food production and preparation of various media and solutions.
Our Media Kitchen and Fly Food staff prepare substantial quantities of reagent quality solutions and media for cell culture, flies (more than 1,700,000 tubes and bottles per year) and other organisms. The Fly Food staff have moved back to the IMP building. There is now more space to create better and more convenient working conditions for preparing fly food and for storing all the goods we need. We also prepare many selected reagents such as DNA molecular weight markers, enzymes, a variety of transformation-competent E. coli strains and we maintain a stock of cloning vectors, sequencing primers and other reagents.
Production of antibodies
The production and isolation of many different monoclonal antibodies in hybridomas in collaboration with IMP group members, and organising antibody production in rabbits with an outside company, takes some of our working time capacity.
Sequencing and DNA isolation
Figure 1 (Click to view legend)
The 48 capillary ABI 3730 DNA Genetic Analyser is the only work horse. The 16 capillary ABI 3100 Genetic Analyser is solely used as a back-up sequencer for emergencies. We sequenced approximately 55,000 samples in the first 9 months of this year. This substantially increased demand is due to screening projects and the new fly library, but also to new groups at IMBA as well as at the IMP. We are primarily using the 3730 DNA Analyser because of its sensitivity and lower consumables running costs. The average read-length is 700–900 bases for standard DNA samples with 50 cm ABI 3730 capillaries. DNA sample quality and concentration are a problem even when prepared by sophisticated Qiagen Kits like Midi-, Maxi- or Minipreps, as are wrong primer sets or insufficiently documented plasmid constructs from outside sources. Compared to analysis by restriction digest, sequencing is faster and easier. The clean-up protocol with Sephadex G50 superfine columns on a 96-well microtiter plate format with optimised Sephadex consistency and centrifugation conditions has been transferred to a BioTek benchtop minirobot. The results tell us that there is no difference between the plates prepared by the robot and those prepared manually and they do not show “dye blobs” with good quality DNA samples.
Selected Publications
2004
Wirth, KG., Ricci, R., Giménez-Abián, JF., Taghybeeglu, S., Kudo, NR., Jochum, W., Vasseur-Cognet, M., Nasmyth, K. (2004). Loss of the anaphase-promoting complex in quiescent cells causes unscheduled hepatocyte proliferation.
Genes Dev. 18(1):88-98
(abstract)
