Description
The DNA sequencing facility developed from the Insect Molecular Genetics Lab (K. Louis' group) in the Drosophila sequencing project. It has been offering service sequencing for the IMBB as well as the wider scientific community of Crete for the past 9 years. Its sequencers are housed in room A020, together with the transcriptomics lab, while the preparation of the samples is carried out in room A030 in K. Louis' laboratory. There is a charge of 10.5 € per reaction for members of the IMBB, while for other sequencing orders the amount charged is agreed upon depending on the number of the reactions. The services are then paid mostly in the form of the direct provision of consumables to be paid for by grants of the lab ordering the sequencing reactions. Funds obtained by the sequencing facility are at the discretion of the IMBB Director.
Equipment
An ABI 377 sequencer which was upgraded into XL is the sole sequencing equipment at this moment. It is considered outdated and there is no support by the manufacturing company (Applied Biosystems).
Services
The facility provides sequencing on plasmids and PCR product templates made with the Big Dye terminator v.3.1 cycle sequencing kit (Applied Biosystems). If needed, it also provides a limited number of primers: T7, T3 and sp6.
Sequencing length can reach about 1000 bases with uneven quality. Sequences are clipped at the end depending on the quality. The results are deposited in folders in backserv.imbb.forth.gr as specified by the senders who are notified by e-mail when the sequence is available.
Details for the preparation of samples for sequencing and submission can be found at the URL http://konops.imbb.forth.gr/cgi-bin/of.pl . Advice concerning improvement of sequencing and problem solving is provided as part of the service.
The facility may participate in sequencing projects under special agreement between any party interested and the scientist in charge (K. Louis).
Personnel
| Name |
Lefteris Spanos |
| Telephone number |
+30-2810-391149, 391021 |
| Email address |
spanos@imbb.forth.gr |
| Room Number |
A130 |
Contact Person
| Name |
Kitsos Louis |
| Telephone number |
+30-2810-391119 |
| Fax Number |
+30-2810-391104 |
| Email address |
louis@imbb.forth.gr |
| Room Number |
A153 |
Publications
• Favia, G. and Louis, C. (1999). Molecular identification of chromosomal forms of Anopheles gambiae sensu stricto. Parassitologia , 41, 115-118.
• Louis, C. (1999). Malaria entomology: can genomics help?. Parassitologia , 41, 489-492.
• Benos, P. V., Louis, C., Sidén-Kiamos, I. , Bolshakov, S., Papagiannakis, G., Spanos, and Glover, D. M. (2000). From Sequence to Chromosome: the tip of the X chromosome of D. melanogaster . Science . 287, 2220-2222.
• Spanos, L., Koutroubas, G., Kotsyfakis, M. and Louis, C. (2000). The complete sequence of the mitochondrial genome of the Medfly, Ceratitis capitata . Insect Mol. Biol . 9, 139-144.
• Adams MD, Bolshakov S , Siden-Kiamos I , and Venter JC. The genome sequence of Drosophila melanogaster . (2000) Science . 287, 2185-95.
• Arcà, B., Lombardo, F., Lanfrancotti, A., Spanos, L., Veneri, M., Louis, C. and Coluzzi, M. (2002) A cluster of four D7-related genes are expressed in the salivary glands of the African malaria vector Anopheles gambiae. Insect Mol. Biol. , 11, 47-56.
• Peter, A., Bolshakov, S., Siden-Kiamos, I., Papagiannakis, G., Spanos, L., Louis. C, and Schafer, U. (2002) Identification and mapping of vital gene functions of the Drosophila X-chromosome . EMBO Rep. 3, 34-38.
• Holt, R.A., Louis, C. and Hoffman, S.L. (2002) The Genome Sequence of the Malaria Mosquito Anopheles gambiae ; Science 298, 129-149.
• Parmakelis, A., Spanos, L., Papagiannakis, G., Louis, C. and Mylonas, M. (2003) Mitochondrial DNA phylogeny and morphological diversity in the genus Mastus (Beck, 1837). A study in a recent (Holocene) island group (Koufonissi, SE Crete). Biol. J. Linnean Soc. , 78, 383-399.
• Mongin, M., Louis, C., Holt, R. A., Birney, E. and Collins, F. H. (2004). The Anopheles gambiae genome: an update on sequence, data, annotation and access. Trends Parasitol. , 20, 49-52 .
• Parmakelis, A., Pfenninger, M., Spanos, L., Papagiannakis, G., Louis, C., and Mylonas, M. (2005) Inference of a radiation in Mastus (Gastropoda, Pulmonata, Enidae) on the island of Crete. Evolution 59, 991-1005.
• Parmakelis, A., Stathi, I. , Spanos, L., Louis, C., Mylonas, M. (2006). Phylogeography of Iurus dufoureius (Brullé, 1832) (Scorpions, Iuridae) in the Aegean Archipelago. J. Biogeogr. 33, 251-260.
• Parmakelis, A., Stathi, I. , Chatzaki, M., Simaiakis, S., Spanos, L., Louis, C. and Mylonas, M. (2006). Inference of the evolutionary history of Mesobuthus gibbosus (Brullé 1832) in the north-eastern Mediterranean region. Molec. Ecol. , 5, 2883-2894
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