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Frequently Asked Questions - Sanger Sequencing
Product & Services
The Barcode Economy Run is Microsynth‘s most demanded standard sequencing service for plasmids and PCR products in 1.5 ml tubes. It offers the best price-performance ratio due to its prepayment structure (minimally 50 barcode labels corresponding to 50 sequencing reactions have to be purchased in advance).
Microsynth’s Economy Run corresponds to the Barcode Economy Run, with the exception that customers will be invoiced after using the service and they are not bound to batches of 50 barcodes.
The Premium Run is Microsynth‘s superior sequencing service in 1.5 ml tubes offering a broad spectrum of additional services (e.g. Reaction conditions are optimized specifically for your sample and special treatment protocols for difficult sequences) and troubleshooting protocols (e.g. alternative design of sequencing primers, one-drop sequencing in case of low DNA amounts).
Generally speaking, the Premium Run is designed for customers requiring the best possible outcome along with superior service.
Please login to our webshop to find a sample drop box in your vicinity.
- Barcode Economy Run or Economy Run Service samples: 4 working days
- Premium Run Samples: 3 months
- High-Throughput sample plates: 1 month
- Your specific sequencing primers will be kept at our sequencing lab for at least 6 months (or for 12 months in case you have added them to your “Custom Primer List”).
On request, Microsynth will ship your specific sequencing primer(s) back to your or any other desired address.
Click here to open our standard primer list with all the primers you can select from.
Please note: Microsynth’s webshop allows you to maintain a customized primer list. In case you frequently use more specific sequencing primers, just send them to Microsynth and update your customized primer list. Since Microsynth operates a well-recognized oligo production facility, you can also outsource design and/or synthesis of your sequencing primers.
Order Related Questions
Dependent on the selected service type, your samples have to fulfill different requirements. For following services, Microsynth provides user guides containing a lot of helpful information:
Please click on the name of the service above to open the relevant user guide; or go to the “Sample Requirements” area where you will also find useful information about sample requirements etc.
- Enter our webshop
- Click on Option&Preferences in the DNA Sequencing area
- Click on Order History - Review/Delete Previous Orders
In general, failed sequencing reactions will be automatically repeated in the following mode: in a first attempt one sample will be repeated in order to evaluate the potential for further improvement by modifying the chemistry of the sequencing reaction. If the repetition yields in a better sequencing result, other samples of your order will be resequenced, too. In addition, it is always possible to ask for specific repetitions of failed reactions (best done by phone or email). Be aware, that we keep Economy and Barcode Economy Run samples only for 4 working days.
Go to the “Sequencing Primers” area where you will find a lot of useful information about sequencing primers.
Microsynth provides the sequencing results in text format (FASTA files) and in chromatogram format (ab1 files). In order to open the ab1 files you need to download one of the software tools which are provided in the Software and Links area.
Direct sequencing via the Sanger approach is possible for genomes < 1Mb. However, whenever feasible, we recommend you to amplify the region of interest by PCR and to send us the PCR product(s) for sequencing.
In order to obtain reliable sequencing results, impurities such as dNTPs, PCR primers etc. must be eliminated. Furthermore, it is important to quantify the purified PCR product. This is easily accomplished by performing agarose gel electrophoresis of your PCR product and comparing the band for your PCR product with standard bands of defined concentrations. We do not recommend quantification approaches based on UV spectrophotometric analysis techniques. For successful sequencing, PCR products must be single-banded on the agarose gel. If this is not the case, you should recover the correct band by gel extraction. Using the Premium Run we can do this for you.
The results of the Sanger sequencing are kept for at least 3 months on the webshop.