DNA Sequencing Guidelines

Please carefully follow these guidelines when preparing samples for sequencing. Also remember that placing an order implies your agreement with our terms and conditions.

1. Send the Template DNA in 200 ul PCR tubes, 8-well strips, or 96-well microplates. Dissolve the DNA in 10 ul Tris (10mM, pH 8.0 to 8.5), which corresponds to the Qiagen EB-Buffer solution. Whenever possible, avoid buffers containing EDTA. Use the following reference final concentrations:

Template:	Template Concentration:
PCR Products < 500 bp	2 ng / ul
PCR Products > 500 bp	10 ng / ul
Plasmids < 8 kb	50 ng / ul
Plasmids > 8 kb	150 ng / ul



2. If needed, add 1 ul Primer solution (10 pmol / ul). You can use your own primers or choose a standard primer to be added by Synergene Biotech (service is free of charge). See our primer list for details.


3. Fill out the submission Excel File. Click button to download:



4. Attach this file to an email message and send it to sequencing@synergene-biotech.com. Please remember to save the file before attaching it, otherwise we will not be able to read it.


5. Send us your samples by A-post (inside Switzerland) or any fast shipping service when ordering from other countries. For your convenience, upon request we provide free addressed padded bags and tubes.

The following tips are drawn from practical experience. They can be helpful whenever commercial kits from suppliers such as Promega, Qiagen, etc. are used to purify DNA templates:

1. Make sure you do not overload the columns with bacteria when extracting plasmid DNA since in such cases, the process of purification tends to be less efficient.
   


2. Both PCR and plasmid purification work best if you do not try to recover the eluate down to the very last drop. Commercial kits yield DNA of excellent quality but overelution can put unwanted dirt back to your sample, which leads to poor sequencing results. Since commercial protocols have been extensively tested, it is recommended to follow them exactly. Please, never reload nor use commercial columns more than once, as this can severely compromise the purity of your sample.
   


3. Please make sure that no EDTA residues are left inside your sample. Some commercial kits use elution buffers containing EDTA, such as TE buffers. EDTA complexes magnesium ions, which reduces the effectiveness of the sequencing polymerase. The results of your sequencing can therefore be severely compromised. Please contact us if your protocols include steps using buffers containing EDTA.
   

The best free trace file viewer is definitely Finch TV by Geospiza Inc. It is available for Microsoft Windows, Mac OS X, Linux, and Solaris. Click the icon to download:

Users of Mac OS 8 and 9: Click here for guidelines.

Please also have a look at our software tools page for more software suggestions.

An explanation of the troubleshooting process, as well as a list of information we might need for successful troubleshooting can be found here.

Should you have any questions do not hesitate to contact us. Requests for support related to DNA sequencing should be sent to sequencing@synergene-biotech.com.