About DNA Sequence Analysis Service

For use of this service, you must get permission from your laboratory advisor and submit an application via Webform. After the application is accepted, please bring your samples.

●Service Outline

DNA Sequence Analysis Service is done three times a week on Monday, Wednesday, and Thursday.

Data is usually sent the day after the analysis date (Tuesday, Thursday, and Friday).
(However, there may be a delay in sending the results due to business reasons.)

ABI3500xl Genetic Analyzer (24 capillary type) is used.
All the operations will be done by the staff of our facility.
Please use Webform for application (accessible only within the university campus area).
Fill in the required information according to the instructions and press the “Confirm” button. After confirming the information, press “Send” button to sent an e-mail to the Management Office.

We will inform you of the scheduled date and time to start the analysis.
Please bring your samples to Gene Research Center Room 301 on the scheduled date and time.
After the analysis is completed, the data will be compressed and sent as an attachment via e-mail.

The order of analysis will be on a first-come, first-served basis. After receiving your application, we will contact you by e-mail. The analysis date cannot be specified.

Please follow the process of “Application → Confirmation of scheduled date → Submission”.
 
 
● Preparation of DNA samples

・Preparation in your own laboratory

At least 20µl of dissolved solution is required for purification by Ethanol precipitation + deionized Formamide dissolution and 20µl of supernatant solution for purification by BigDyeX Terminator. This amount is a recommendation and may evaporate over time, so please submit a generous volume.

In the case of the method using Ethanol precipitation + Formamide dissolution, a decrease in fluorescence intensity is observed about 2 days of dissolution. In the case of BigDyeX Terminator purification method, it can be stored at room temperature for 2days or refrigerated for 10days. In either case, store in a darkened container such as aluminum foil wrapped case.

・Bring the sample tubes to Room 301

1. Write the sample name on the tube, transfer the sample tube to the special case provided by the facility, and cover it.

*To prevent mistakes in naming the tubes, please write the sample names so that they can be distinguished from other people’s samples. Also, sequential numbers should be placed at the end.
(ex. If naming only 1,2,3…, it is difficult to distinguish from other user.)

2. Put a sticky note with the date, name, and number of samples on the cover of the special case, and put it in the designated place in the refrigerator of Room 301.


 
 
● About Data

We have received a few e-mails from users after we have sent the results of sequencing analysis, asking us to send the results again because the seq file is missing.
Usually, if there is no seq file in the results, it is because the analysis results were not good and the data could not be handled as a sequence.
In such cases, it may help you to clarify the cause by displaying display the chromatogram (waveform) file using appropriate analysis software.

The Gene Research Center website has a page titled “13 Experiments and Methods of Use,” which includes an article titled “About DNA sequence waveform data editing software.
The information on software for MAC OS and Windows is listed, so please use the appropriate software for your PC environment.
If you are using Windows, Sequence Scanner Software by Lifetechnologies is easy to use. This software allows you to reference waveform data from ab1 file (we have confirmed that it works on Windows 7 and Windows 10 in addition to Windows XP).
Sequence Scanner Software can also be installed on MACs. For details, please refer to “Sequence Scanner/Peak Scanner for Mac Installation Manual 2_PlayOnMac_kagoshima-u ver.“.
If you have any questions about chromatography data, please feel free to contact us.
 
 
● DNA Sequencing Fees, etc.
DNA sequencing fee: Please refer to “here“.
Consumables fee: Please refer to “here“.

 Sequencing kit BigDye Terminator v3.1 (Life technologies)
*One tube of ×5 Sequence buffer(75µl) is included(second ×5 Sequence buffer tube is available for a fee).
 Hi-Di Formamide(500µL)
 Purification kit BigDyeX Terminator (Life technologies)

All fees are for intramural users. For more details, please contact the Management Office.
 
● Information related to DNA sequencing
Useful documents and information about DNA sequencing are available on our website. Please make use of them.

 Please refer to “here“.

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