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BecA Hub in action The hub Service units SegoliP | SegoliP unit |
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Appolinaire Djikeng, email: This e-mail address is being protected from spam bots, you need JavaScript enabled to view it Laboratory / Research Technologists: Ben Kiawa, Lucy Muthui, Inosters Nzuki The SegoliP unit was established in 1993 as a centralized sequencing unit for the International Livestock Research Institute (BecA). In 2004, the unit’s services expanded to include DNA Fragment Analysis support for Genomic Research with the acquisition of the ABI 3130 and ABI 3730 Genetic Analyzers. SegoliP unit Services: The SegoliP unit offers the following services to researchers at the BecA Hub and to institutions in the region:
 DNA Sequencing services Sequencing at the SegoliP unit is primarily conducted with BigDye™ Terminator technology. The sequencing products are run on either the ABI 3730 or ABI 3130 Genetic Analyzers. Ub addition, we have just established the massively parallel sequencing -by-synthesis using the Roche/454 pyrosequencer. Sequencing results:
Requirements for Sequencing: Please supply DNA templates at the following amounts/concentrations:
Please ensure that your DNA templates are highly purified before submitting to SegoliP for sequencing. We recommend the following DNA purification kits: Plasmids
Please ensure that your plasmid templates contain insert before submitting to SegoliP. PCR Products
Please ensure that your PCR products are a single band when analysed by gel electrophoresis before submitting to SegoliP. PCR products with more than one band frequently produce poor results. PCR products sequenced with the PCR primers often give acceptable results. However, for some PCR product templates, it is necessary to sequence with a nested primer or a primer with a 5’ GC clamp, for best results. Sequencing primers When sequencing from pUC, Bluescript and pGEM plasmids, customers are not required to supply forward and reverse vector primers as these are provided by SegoliP. All other sequencing primers must be supplied by the customer. For single sequencing reactions, please supply 10 μl of 5 pmol/μl of each primer. Sequencing primers should have a Tm of 50-58°C. Fragment Analysis services The unit accepts multi-well plates containing DNA fragments for size analysis on the ABI Genetic Analyzers. The instruments use the G5 filter set; four dyes can therefore be used to label PCR fragments: 6-FAM (blue), VIC (green), NED (yellow), PET (red), and LIZ dye is used as the internal size standard. Data is posted on the SegoliP server as it is generated. Customers can access the information at their convenience. The unit also offers eight computers with GeneMapper software for Microsatellite analysis, Basic Sizing/Peak Detection, LOH.  Consumable services A limited number of consumables are available from SegoliP on a cost recovery basis. These include 96-well and 384-well reaction plates, GeneScan 500 LIZ Size Standard and Hi-Di Formamide. Oligonucleotide Synthesis services Each month, the SegoliP unit sends a consolidated order to an overseas company for oligonucleotide synthesis. From order to receipt of the oligos is usually about 10 days. Oligos are transported in a lyophilised form, HPSF (High Purity Salt Free), and come with full documentation that includes the oligo name, sequence, amount of oligo, molecular weight, melting temperature and GC content. For further details on the modifications and scales of synthesis of oligos, please visit the website of the company we use: http://www.bioneer.com Cost of Services Sequencing
Fragment Analysis
Oligonucleotide Synthesis
With overhead and shared freight cost included, the total cost of oligos is normally about US$ 1 per base for 0.01 μmol scale.   Download the SegoliP unit brochure  SegoliP Unit Brochure
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Thu, 22 July 2010,
04:38:50 AM
Thu, 22 July 2010,
04:38:50 AM
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