Ready-to-Load Sequencing (SS1005)
For Ready-to-Load samples, customers undertake the first two steps in Automated Sanger Sequencing; Cycle-sequencing and Clean-up, and only send us the dried precipitate after Clean-up. We will perform Loading and Data Analysis.
We accept reactions performed with ABI® PRISM BigDye Terminator Cycle Sequencing Kit v3.1. If you are using other sequencing chemistry kit, please acquire from your local Sales Office.
Results may be expected in 1 - 2 working days upon order acknowledgement.
Both cycle sequencing reaction setup and post-cycle sequencing clean-up are equally crucial for quality of DNA sequencing results. Poor handling in either step will compromise the result significantly.
About your setup of cycle sequencing reaction
Many of our Ready-to-load customers have modified or optimised their sequencing reactions with a novel protocol, for example diluting BigDye® sequencing mix, using an increased number of cycles, adding DMSO and etc. We may not be able to provide support for reactions that do not adhere to our recommended guidelines.
About your post-cycle sequencing clean-up
We recommend fresh BigDye® reactions to be purified immediately. If this cannot be done, take them out from your thermocycler and freeze them at -20 deg C over the weekend.
You may use the standard EDTA-Sodium Acetate-Ethanol Precipitation (which is recommended by Applied Biosystems for BigDye® Terminator Cycle Sequencing Kit v3.1). This requires skilful handling. Other clean-up methods in the market for post-sequencing include column/ plate purification using gel bed filtration, magnetic beads, ethanol precipitation with co-precipitant and etc. As long as there is no carry-over of ethanol residue or water after the drying process, your samples should be fine and Ready to Load. For reaction(s) purified by magnetic beads, please indicate the elution volume (by water).
You are not required to add deionised formamide or Hi-Di® formamide on your Ready to Load samples. We will add formamide and perform the denaturation on our thermocyclers prior loading.
If required, we can also help to troubleshoot the failure of customers’ cycle sequencing reaction setup and post-cycle sequencing clean-up. Please contact your Local Sales Office for assistance. Charges apply. You can send us the following: -
- the DNA template and its sequencing primers in separate tubes (as in our service SS1001);
- Pre-cycle reactions, which is without post-cycle sequencing clean-up(as in our service SS1019); and
- Ready to Load samples, where the customer performs the cycle sequencing reactions and post-cycle sequencing clean-up at their own (as in our service SS1005).