Brief Information
The SouthPath & Flinders Sequencing Facility provides a sequencing and genotyping service to internal university staff and external clients. We currently use an Applied Biosystems 3130xl Genetic Analyser with BigDye Terminator v3.1 chemistry.
Sequencing
The service includes
- cycle sequencing using BigDye Terminator v3.1 chemistry.
- Analysis of products using ABI 3130xl Genetic Analyser.
- Examination of chromatograms including basic editing of raw data.
- Universal primers included at no additional cost. These include T3, T7, M13 forward and reverse, SK, KS and SP6.
- Turnaround time of 3-5 working days.
The amount of DNA that is required
Sample Type | Volume Required | Concentration Required |
Double- and Single-stranded DNA | 10µl | 100ng/µl |
Double-stranded PCR product | 10µl | 10ng/µl per 100 bp of Template (i.e. 30ng/µl for 300 bp product) |
Primers | 2µl | 5µM |
Genotyping
Genotyping samples should be provided as fluorescently labelled PCR products. The concentration of products needs to be optimised by the submitter prior to submission.
The service includes appropriate internal size standards. The instrumentation supports the Virtual Filter Sets D, E5 G5 :
| Filter Set | Blue | Green | Yellow | Red | Orange |
| D | 6-FAM | HEX | NED | ROX | |
| E5 | dR110 | dR6G | dTAMRA | dROX | LIZ |
| G5 | 6-FAM | VIC | NED | PET | LIZ |
| F | 5-FAM | JOE | NED | ROX |
Filter Set E5 can be used to run SNaPshot multiplex reactions.
We are equipped to process 96- and 384-well plates.
Costs
Sequencing costs:
$12.75 - per template. DNA supplied by customer
$6 - per template, electrophoresis only. Sequencing reaction supplied by customer
$0.50 - colour chromatograms
Genescan costs:
$4.80 - per analysis
Discounts apply to Flinders Medical Centre and Flinders University customers.
Results
Turnaround time is usually 3-5 working days but is dependant on demand. Results can be sent electronically via email or ftp, by mail or put on to disk for an additional $1.
Request Form
A Microsoft Excel-based electronic request form for both sequencing and fragment analysis/genotyping (XLS 309KB) is available for download. Please email the form to the Facility when sending samples.
Please address samples to :
Sequencing Facility
Dept. of Haematology
Flinders Medical Centre
Email : dna.seq@flinders.edu.au
Further Information
For further information, please contact:
Oliver Van Wageningen.
TEL: 08 8204 5097
Scott Grist
TEL: 08 8204 5275
Email: dna.seq@
