Sanger Sequencing Repeat Policy
Troubleshooting Terms
The total cost of the order is dependent on the number of reactions performed by our technologists in the initial run, regardless of the quality of data returned. Any repeated reactions are performed as a courtesy and may be selected for re-sequencing based upon the analyst’s judgement when reviewing the results of the initial run. There is no additional charge for repeated reactions.
The first 60-70 bps of a sequence may be noisy or unreliable, as that region is directly after the primer binding site. ACGT only guarantees up to 750 bps of usable data for plasmids. If you are having trouble with sections of the sequence outside of the range of 60-750 bps, you may need to use additional primers for accurate results.
General
In the event that all reactions using the same template or primer fail, those reactions may not be repeated. The specific template or primer will be detailed in the results summary.
Reactions producing data with double peaks, mixed PCR sequences, repetitive sequences, or homopolymers will not be repeated. These outcomes are typically not a result of sequencing reaction failure, but are caused by the sequence itself or the primer used.
Reactions producing data with clear evidence of high GC content, hairpins, or other secondary structures will not be repeated with our standard reaction protocol. If hard to sequence regions are observed and noted as such in the results summary, it is recommended to place a new order for those reactions, with either our Hairpin or High GC protocol selected. Additional charges may apply.
Low Cost Option (LCO)
Troubleshooting will be performed on a subset of reactions that do not meet our quality standards after the initial run. Depending on troubleshooting results, the remainder of the order may or may not be repeated. High intensity data will not be diluted and repeated, as samples should be premixed to the concentrations specified in our guidelines.
Standard
Troubleshooting will be performed on a subset of reactions that do not meet our quality standards after the initial run. Depending on troubleshooting results, the remainder of the order may or may not be repeated. PCR products may be diluted in the event that the results show evidence of high intensity data.
Premium and Direct Colony Service
In most cases, if the initial reaction does not produce usable data (i.e. bad or noisy data), ACGT will repeat the reaction, including dilutions of high intensity data for both PCR products and plasmids. For higher volume orders and orders where quality checks reveal issues with sample quality, troubleshooting reactions may be performed and based on the result, the remainder of the order may or may not be repeated.