Sequencing FAQs

Full sequencing service– you provide the template and primer mix, we do the sequencing reaction, clean up and capillary separation.

Clean-up and separation only– you do the sequencing reaction, we do the clean up and capillary seperation. 

You can view our sequencing reaction set up guide in the Sequencing Submission Portal to help you with setting up your Sequencing reaction.

Full sequencing service– Samples are to be submitted in clearly labelled 1.5mL tubes. Each sample typically comprises a template and single primer mix made up to a total volume of 15.5ul (where necessary, add water to make up the remaining volume). Please refer to our sequencing reaction set up guide in the Sequencing Submission Portal for the amount of template and primer to be added.

Clean-up and separation only–  Samples (20µl sequencing reaction) submitted in clearly labelled 0.2mL tubes/strips. Our sequencing reaction set up guide in the Sequencing Submission Portal has suggested reaction volumes and cycling conditions.  Most people start with using 1µl of Big Dye and move to 0.5µl or less once sequencing is optimised.

Depending on the type of product you are trying to sequence a guideline for the recommended amounts can be seen in the below table;

Primer: 3.2pmol

We strongly recommend running a small amount of your DNA (either PCR product or plasmid) on a gel to quanitfy the amount of DNA template.

If you already have an account with us you just need to fill in your sequencing submission form with the sample details and email it to us at uqgrs@uq.edu.au prior to dropping off your samples.

Samples can then be submitted in person to Genetic Research Services prior to 12pm for processing that day. Samples submitted after 12pm will be processed the following work day.

Alternatively, samples can be submitted via the internal mail or sent via Australia Post.

If you or your research group does not have an up to date account with us a RISQ form will also need to be filled out in order to create one. Contact us or log in to the Sequencing Submission Portal to obtain a new RISQ form.

If submitted before 12pm results will be available by close of business the working day following sample submission for samples submitted for clean up and separation. Results for samples submitted for full sequencing will be available by close of business two working days following sample submission.

A priority service is also available, contact us to enquire about this service.

All result reports are saved on a restricted-access shared drive. Only individuals with approved access will be able to view the results. Results will be made available on the shared drive for a month. 

Access to your results folder can be authorised by Genetic Research Services with the provision of your details and research group.

If you are unable to access the shared drive results will be emailed directly.

Results are reported as FSA files and may be viewed in a free downloadable software of your choice. Genetic Research Services recommends Peak scanner or Finch TV to view your sequencing results.

You will receive a 10% discount for FULL plate submissions – 95 samples for our clean and separation service and 94 samples for our full sequencing service.

For large batches of samples (greater than 32 samples), please submit samples in 96-well plates. For clean-up and separation only samples, please leave one well empty for our internal control. For full sequencing 2 internal controls are used (1 for the full sequencing reaction and 1 for the clean-up and separation reaction) so 2 wells will need to be left empty on these plates.

Genetic Research Services sell the BigDye sequencing reagents in small aliquots if you are interested in doing the reaction yourself without the outlay of a full kit. Otherwise kits can be purchased direct from ThermoFisher.

Sequencing may not work as expected for a variety of reasons. The first things to check when your sequencing results are not as expected are the quality and concentration of both your primers and template. Other reasons may include salt contamination, the size of the fragment you are trying to sequence and the quality of your clean up process.

Hairpin or G-C rich regions can also be particularly troublesome for sequencing. In these instances the use of BDT V1.1 is recommended in your sequencing reaction.

If you are still having troubles with sequencing submitted to us contact us with the names of the samples and the date they were submitted and we will work with you to try and help you resolve your issues.